Treatment of Epithelial Cysts by Intracystic Injection of Antineoplastic Particles

ABSTRACT

Disclosed herein are methods for treating epithelial cysts, including pancreatic cysts, in a subject by intracystic injection of compositions comprising antineoplastic particles, including taxane particles such as paclitaxel particles and docetaxel particles.

CROSS REFERENCE

This application claims priority to U.S. Provisional Patent Application Ser. No. 62/617,711 filed Jun. 9, 2017, incorporated by reference herein in its entirety.

FIELD OF THE INVENTION

The present invention generally relates to treatment of epithelial cystic neoplasms (cysts).

BACKGROUND

A cystic neoplasm (cyst) is an abnormal sac in the body that may be filled with a liquid or semisolid substance. A cyst can form in any part of the body, including bones, organs and soft tissues. Generally, cysts are noncancerous (benign), however, a benign cyst can be precancerous and change into a malignant growth. Epithelial cysts are cysts which have an inner epithelial lining. Examples of epithelial cysts include gastrointestinal cysts such as hepatic cysts, pancreatic cysts, splenic cysts, colon cysts; urologic cysts such as renal cysts, epididymal cysts, prostatic cysts; gynecological cysts such as ovarian cysts and vaginal cysts; head and neck cysts such as thyroid cysts, parathyroid cysts, and other head and neck cysts; as well as other cysts such as Baker's cysts, lung cysts, lymphatic cysts, and pericardial cysts.

Pancreatic cystic neoplasms are being detected with increasing frequency due to improved cross-sectional imaging and routine examination (Pitman 2010). Mucinous pancreatic cystic neoplasms represent approximately 75% of all pancreatic cystic neoplasms and are divided into two categories: intraductal papillary mucinous neoplasms (IPMNs) and mucinous cystic neoplasms (MCNs). Both types of mucinous pancreatic neoplasms may present with symptoms such as abdominal pain, pancreatitis, jaundice, weight loss, malabsorption, nausea, vomiting, and palpable abdominal mass (Tanaka 2005; Muthusamy 2016). Oftentimes, the cysts do not present any symptoms at all and are detected by chance because of the improved imaging when patients are being examined for other reasons.

Mucinous pancreatic cystic neoplasms have malignant potential, thus each patient diagnosed is at risk for developing pancreatic cancer (Farrell 2015; Sarr 2000). In a study of 401 patients with pancreatic cysts, 11% of resected cysts contained invasive cancer (Ferrone 2009). Patients diagnosed with IPMN have a 40% chance of invasive cancer and 58% have underlying malignant features, while those with MCN have a malignancy risk that varies between 10% and 50% (Greer 2016; Allen 2007). To avoid progression to pancreatic cancer, an aggressive and rapidly fatal disease, all patients with mucinous pancreatic cysts should be carefully evaluated and offered treatment (National Cancer Institute 2016; Fernández-del Castillo 1995).

An emerging treatment for mucinous pancreatic cysts is cyst fluid aspiration by endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) followed by endoscopic ultrasound-guided (EUS-guided) ethanol lavage as an ablative agent in order to induce cell death by membrane lysis, protein denaturation, and vascular occlusion (Jani 2011; DeWitt 2009). However, studies examining cyst volume response to ethanol injection failed to standardize their techniques and ethanol concentration/volume, and a ten-year study conducted by Gómez 2016 demonstrated that only 9% of patients experienced complete cyst resolution (Gomez 2016; Kirtane 2016). There has been same controversy regarding the current treatment methods and practices for pancreatic cysts (Farrell 2013, Tanaka 2016, McGrath 2017).

Current methods for determining malignancy within mucinous cysts provide unreliable results. It is difficult to identify cyst histology without resection. Approximately 25% of non-operative histologic diagnoses are inaccurate. Physicians compensate for this deficit by recommending that patients with cysts with features such as symptoms, positive cytology, mural nodules, or greater than 3 cm undergo surgical resection (Pitman 2010). Due to the concerns of mucin leakage, pancreatic fistulae, and recurrence, the standard surgical treatment for invasive and non-invasive MCNs and IPMNs is pancreatectomy with lymph node dissection, rather than focal non-anatomic resections or resections without lymphadenectomy or splenectomy (Tanaka 2012). In a study of 37 high-volume centers (2694 patients), the pancreatectomy mortality rate was reported to be 1.3%, increasing to 3.0% in patients 80 years of age and over, with overall complication rates ranging from 20.0% to 72.2% (Tamirisa 2016). In patients that can be diagnosed with a malignancy rate of less than 3%, the risk of death from pancreatectomy is higher than that of malignancy (Allen 2007). Even after surgery, the recurrence rates of cysts can be as high as 20% (Tanaka 2012). Untreated cysts, however, may progress to malignant disease while under observation (Allen 2007).

SUMMARY OF THE INVENTION

The present invention provides solutions to the aforementioned limitations and deficiencies in the art relating to treatment of cystic neoplasms (cysts), particularly to treatment of epithelial cysts including pancreatic cysts, by injection of antineoplastic particles, such as taxane particles, directly into the cysts (intracystic injection). When a composition of antineoplastic particles, such as taxane particles (e.g., paclitaxel particles or docetaxel particles), are injected into a cyst, the antineoplastic particles will persist in the cyst for a longer time than a solution of antineoplastic agent injected into the cyst because solutions are more easily and quickly cleared from the cysts by the body. Although not bound by theory, it is hypothesized that the use of antineoplastic particles result in increased efficacy and decreased toxicity as compared with solutions of antineoplastic agents, at least in part, due to the slow release of the antineoplastic agent from the suspended particle. It also is hypothesized that because of the physical characteristics of the antineoplastic particles, they can embed within or on the inner epithelial lining of the cyst resulting in a longer residence time than with solutions or with albumin coated particles. Instillation of a suspension of antineoplastic particles directly into the pancreatic cyst creates a depot effect where the antineoplastic agent is slowly released from the particles into the cyst, resulting in prolonged local exposure of the antineoplastic agent. Clearance from the pancreas is reduced, with lower systemic levels of antineoplastic agent, further limiting systemic toxicity. In a preferred embodiment, the antineoplastic particles are taxane particles such as paclitaxel particles or docetaxel particles.

In one aspect of the invention, disclosed are methods for treating epithelial cysts, the methods comprise injecting a composition comprising an effective amount of antineoplastic particles directly into the cyst (intracystic injection) thereby treating the epithelial cyst, wherein the particles have a mean particles size (number) of from 0.1 microns to 5 microns. In preferred embodiments, the antineoplastic particles are taxane particles. In some embodiments, the particles of the antineoplastic agent have a mean particle size (number) of from 0.1 microns to 1.5 microns. In some embodiments, the taxane particles are paclitaxel particles. In some embodiments, the composition and taxane particles exclude albumin. In other embodiments, the taxane particles are docetaxel particles. In some embodiments, the epithelial cyst is a pancreatic cyst such as a mucinous pancreatic cyst. In some embodiments, the epithelial cyst is reduced in volume/size, has reduced growth rate, is eliminated, or is ablated after treatment by injection of the composition. In some embodiments, the pain associated with an epithelial cyst is reduced after injection of the composition.

In another aspect of the invention, disclosed are kits comprising: (a) a first vial comprising taxane particles; (b) a second vial comprising a pharmaceutically acceptable aqueous carrier and a surfactant; and (c) instructions for reconstituting the antineoplastic particles into a suspension useful for intracystic injection by: combining the contents of the first vial and the second vial to form the suspension and optionally diluting the suspension with a diluent. In some embodiments, the taxane particles have a mean particle size (number) of from 0.1 microns to 1.5 microns. In some embodiments, the surfactant is a polysorbate. In some embodiments, the taxane particles are paclitaxel particles. In other embodiments, the taxane particles are docetaxel particles.

Also, disclosed in the context of the present invention are the following embodiments 1 to 39:

Embodiment 1 is a method for treating an epithelial cyst, the method comprising injecting a composition comprising an effective amount of antineoplastic particles directly into the cyst, thereby treating the epithelial cyst, wherein the particles have a mean particle size (number) of from 0.1 microns to 5 microns. Embodiment 2 is the method of embodiment 1, wherein the composition further comprises a liquid carrier, and wherein the composition comprises a suspension of the antineoplastic particles dispersed in the carrier. Embodiment 3 is the method of embodiment 2, wherein the carrier is an aqueous carrier. Embodiment 4 is the method of embodiment 3, wherein the aqueous carrier comprises 0.9% saline solution. Embodiment 5 is the method of any one of embodiments 3 or 4, wherein the aqueous carrier comprises a surfactant. Embodiment 6 is the method of embodiment 5, wherein the surfactant is a polysorbate. Embodiment 7 is the method of embodiment 6, wherein the polysorbate is polysorbate 80, and wherein the polysorbate 80 is present in the aqueous carrier at a concentration of between about 0.01% v/v and about 1% v/v. Embodiment 8 is the method of any one of embodiments 3 to 7, wherein the composition further comprises a diluent, wherein the carrier and the diluent form a mixture, and wherein the composition is a suspension of the antineoplastic particles dispersed in the carrier/diluent mixture. Embodiment 9 is the method of embodiment 8, wherein the diluent is a 0.9% saline solution. Embodiment 10 is the method of any one of embodiment 1 to 9, wherein the antineoplastic particles are taxane particles. Embodiment 11 is the method of embodiment 10, wherein the taxane particles comprise at least 95% of the taxane, wherein the taxane particles have a mean particle size (number) of from 0.1 microns to 1.5 microns, and wherein the composition and taxane particles exclude albumin. Embodiment 12 is the method of any one of embodiments 10 to 11, wherein the concentration of taxane particles in the composition is between about 6 mg/mL and about 15 mg/mL. Embodiment 13 is the method of any one of embodiments 10 to 12, wherein the taxane particles are paclitaxel particles. Embodiment 14 is the method of embodiment 13, wherein the paclitaxel particles have a specific surface area (SSA) of at least 18 m²/g, 20 m²/g, 25 m²/g, 30 m²/g, 32 m²/g, 34 m²/g, or 35 m²/g; or from about 18 m²/g to about 50 m²/g. Embodiment 15 is the method of any one of embodiments 13 to 14, wherein the paclitaxel particles have a bulk density (not-tapped) of 0.05 g/cm³ to 0.15 g/cm³. Embodiment 16 is the method of any one of embodiments 10 to 12, wherein the taxane particles are docetaxel particles. Embodiment 17 is the method of embodiment 16, wherein the docetaxel particles have a specific surface area (SSA) of at least 18 m²/g, 20 m²/g, 25 m²/g, 30 m²/g, 35 m²/g, 40 m²/g, or 42 m²/g; or from about 18 m²/g and about 60 m²/g. Embodiment 18 is the method of any one of embodiments 16 to 17, wherein the docetaxel particles have a bulk density (not-tapped) of 0.05 g/cm³ to 0.15 g/cm³. Embodiment 19 is the method of any one of embodiments 1 to 18, wherein the cyst fluid is withdrawn from the cyst prior to injecting the composition. Embodiment 20 is the method of embodiment 19, wherein the volume of the composition injected into the cyst is equal to the volume of cyst fluid removed from the cyst. Embodiment 21 is the method of any one of embodiments 1 to 20, wherein the epithelial cyst is a pancreatic cyst. Embodiment 22 is the method of embodiment 21, wherein the pancreatic cyst is a mucinous pancreatic cyst. Embodiment 23 is the method of any one of embodiments 1 to 22, wherein the injection of the composition is conducted by endoscopic ultrasound-guided fine needle injection (EUS-FNI). Embodiment 24 is the method of any one of embodiments 1 to 25, wherein the epithelial cyst is reduced in volume/size, has reduced growth rate, is eliminated, or is ablated after injection of the composition, and/or wherein the pain associated with the cyst is reduced. Embodiment 25 is the method of any one of embodiments 1 to 24, wherein the epithelial cyst is benign. Embodiment 26 is a kit comprising:

-   -   (a) a first vial comprising taxane particles;     -   (b) a second vial comprising a pharmaceutically acceptable         aqueous carrier and a surfactant; and     -   (c) instructions for reconstituting the taxane particles into a         suspension useful for intracystic injection by: combining the         contents of the first vial and the second vial to form the         suspension and optionally diluting the suspension with a         diluent.         Embodiment 27 is the kit of embodiment 26, wherein the aqueous         carrier is 0.9% saline solution, wherein the surfactant is a         polysorbate, and wherein the polysorbate is at a concentration         of between about 0.01% v/v and about 1% v/v.         Embodiment 28 is the kit of any one of embodiments 26 to 27,         wherein the diluent is 0.9% saline solution.         Embodiment 29 is the kit of any one of embodiments 26 to 28,         wherein the taxane particles comprise at least 95% of the         taxane, wherein the taxane particles have a mean particle size         (number) of from 0.1 microns to 1.5 microns, and wherein the         taxane particles exclude albumin.         Embodiment 30 is the kit of any one of embodiments 26 to 29,         wherein the taxane particles are solid, uncoated (neat)         individual particles; wherein the taxane particles are not bound         to any substance; wherein no substances are absorbed or adsorbed         onto the surface of the taxane particles; wherein the taxane         particles are not encapsulated, contained, enclosed or embedded         within any substance; wherein the taxane particles are not         coated with any substance; wherein the taxane particles are not         microemulsions, nanoemulsions, microspheres, or liposomes;         wherein the taxane particles are not bound to, encapsulated in,         or coated with a monomer, a polymer (or biocompatible polymer),         a protein, a surfactant, or albumin; and/or wherein a monomer, a         polymer or biocompatible polymer), a protein, a surfactant, or         albumin is not absorbed or adsorbed onto the surface of the         taxane particles.         Embodiment 31 is the kit of any one of embodiments 26 to 30,         wherein the taxane particles are paclitaxel particles.         Embodiment 32 is the kit of embodiment 31, wherein the         paclitaxel particles have a specific surface area (SSA) of at         least 18 m²/g, 20 m²/g, 25 m²/g, 30 m²/g, 32 m²/g, 34 m²/g, or         35 m²/g; or from about 18 m²/g to about 50 m²/g.         Embodiment 33 is the kit of any one of embodiments 31 to 32,         wherein the paclitaxel particles have a bulk density         (not-tapped) of 0.05 g/cm³ to 0.15 g/cm³.         Embodiment 34 is the kit of any one of embodiments 26 to 30,         wherein the taxane particles are docetaxel particles.         Embodiment 35 is the kit of embodiment 34, wherein the docetaxel         particles have a specific surface area (SSA) of at least 18         m²/g, 20 m²/g, 25 m²/g, 30 m²/g, 35 m²/g, 40 m²/g, or 42 m²/g;         or from about 18 m²/g and about 60 m²/g.         Embodiment 36 is the kit of any one of embodiments 34 to 35,         wherein the docetaxel particles have a bulk density (not-tapped)         of 0.05 g/cm³ to 0.15 g/cm³.         Embodiment 37 is a method of administering a composition         comprising antineoplastic particles to an epithelial cyst of a         subject, the method comprising injecting the composition into         the cyst using endoscopic ultrasound guided-fine needle         injection, wherein the antineoplastic particles have a mean         particle size (number) of from 0.1 microns to 5 microns, and         wherein the antineoplastic particles are crystalline particles.         Embodiment 38 is the method of embodiment 37, wherein the         antineoplastic particles have a mean particle size (number) of         from 0.3 microns to 5 microns.         Embodiment 39 is the method of any one of embodiments 37 to 38,         wherein the antineoplastic particles are taxane particles.

The term “antineoplastic agents” as used herein are drugs used to treat neoplasms including non-cancerous neoplasms and malignant neoplasms, and include “chemotherapeutic agents”, which are drugs used to treat cancer. In a preferred embodiment, the antineoplastic agent is a taxane.

The terms “antineoplastic agent particles”, “antineoplastic particles” or “particles of an antineoplastic agent”, as used herein are particles of an antineoplastic agent and have a mean particle size (number) of from about 0.1 microns to about 5 microns (about 100 nm to about 5000 nm) in diameter. In a preferred embodiment, the antineoplastic particles are taxane particles.

As used herein, the terms “cystic neoplasm” and “cyst” can be used interchangeably and mean an abnormal sac in the body that may be filled with a liquid or semisolid substance. An “epithelial” cystic neoplasm or “epithelial” cyst, has an epithelial inner lining. In some embodiments, the epithelial cyst is benign. In some embodiments, the cyst is precancerous.

As used herein, the terms “treat”, “treatment”, “treated”, or “treating” with respect to cystic neoplasms (cysts) means accomplishing one or more of the following: (a) reducing cyst volume/size; (b) reducing cyst growth rate; (c) eliminating a cyst; (d) ablation of a cyst; or (e) reducing pain associated with the cyst.

As used herein, the term “suspension” means a suspension dosage form composition where antineoplastic particles are dispersed (suspended) within a continuous carrier or a continuous carrier/diluent mixture. The antineoplastic particles can be completely dispersed, partially dispersed and partially dissolved, but not completely dissolved in the carrier or carrier/diluent mixture.

The terms “subject” or “patient” as used herein mean a vertebrate animal. In some embodiments, the vertebrate animal can be a mammal. In some embodiments, the mammal can be a primate, including a human.

The term “room temperature” (RT) as used herein, means 20-25° C.

The term “surfactant” or “surface active agent” as used herein, means a compound or a material or a substance that exhibits the ability to lower the surface tension of water or to reduce the interfacial tension between two immiscible substances.

As used herein, the singular forms “a”, “an” and “the” include plural referents unless the context clearly dictates otherwise. “And” as used herein is interchangeably used with “or” unless expressly stated otherwise.

The terms “about” or “approximately” as used herein mean+/−five percent (5%) of the recited unit of measure.

For this application, a number value with one or more decimal places can be rounded to the nearest whole number using standard rounding guidelines, i.e. round up if the number being rounded is 5, 6, 7, 8, or 9; and round down if the number being rounded is 0, 1, 2, 3, or 4. For example, 3.7 can be rounded to 4.

Unless the context clearly requires otherwise, throughout the description and the claims, the words “comprise”, “comprising”, and the like are to be construed in an inclusive or open-ended sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of “including, but not limited to”. Words using the singular or plural number also include the plural and singular number, respectively. Additionally, the words “herein,” “above,” and “below” and words of similar import, when used in this application, shall refer to this application as a whole and not to any particular portions of the application. The compositions and methods for their use can “comprise,” “consist essentially of,” or “consist of” any of the ingredients or steps disclosed throughout the specification. With respect to the phrase “consisting essentially of,” a basic and novel property of the methods of the present invention are their ability to treat epithelial cystic neoplasms by intracystic injection of compositions of antineoplastic particles.

It is contemplated that any embodiment discussed in this specification can be implemented with respect to any method or composition of the invention, and vice versa. Furthermore, compositions of the invention can be used to achieve methods of the invention.

The description of embodiments of the disclosure is not intended to be exhaustive or to limit the disclosure to the precise form disclosed. While the specific embodiments of, and examples for, the disclosure are described herein for illustrative purposes, various equivalent modifications are possible within the scope of the disclosure, as those skilled in the relevant art will recognize.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a graph of the flux of paclitaxel (delivered dose of paclitaxel active drug across a porcine bladder membrane over time) from various paclitaxel formulations.

FIG. 2 is a graph of the flux of paclitaxel (delivered dose of paclitaxel active drug across a porcine intestinal membrane over time) from various paclitaxel formulations.

FIG. 3 is a graph of the flux of docetaxel (delivered dose of docetaxel active drug across a porcine bladder membrane over time) from various docetaxel formulations.

DETAILED DESCRIPTION OF THE INVENTION

In one aspect of the invention, disclosed are methods for treating epithelial cysts, the methods comprise injecting a composition comprising an effective amount of antineoplastic particles directly into the cyst (intracystic injection), thereby treating the cyst. In some embodiments, the composition further comprises a carrier, which can be an aqueous liquid carrier. In some embodiments, the carrier comprises a surfactant which can be a polysorbate, such as polysorbate 80. In other embodiments, the composition further comprises a carrier and a diluent, wherein the carrier and diluent form a mixture. In some embodiments, the composition is a suspension wherein the antineoplastic particles are suspended in the carrier or the carrier/diluent mixture. In preferred embodiments, the antineoplastic particles are taxane particles. In some embodiments, the particles of the antineoplastic agent have a mean particle size (number) of from 0.1 microns to 1.5 microns. In some embodiments, the taxane particles are paclitaxel particles. In other embodiments, the taxane particles are docetaxel particles. In various embodiments, the methods further comprise withdrawal of the cyst fluid from the cyst prior to injecting the composition and injecting a volume of the composition equal to the volume of the withdrawn cyst fluid. In some embodiments, the epithelial cyst is a pancreatic cyst such as a mucinous pancreatic cyst. In some embodiments, the injection of the composition is conducted by endoscopic ultrasound-guided fine needle injection (EUS-FNI). In some embodiments, the epithelial cyst is reduced in volume/size, has reduced growth rate, is eliminated, or is ablated after treatment by injection of the composition. In some embodiments, the pain associated with the epithelial cyst is reduced.

In another aspect of the invention, disclosed are kits comprising: (a) a first vial comprising taxane particles; (b) a second vial comprising a pharmaceutically acceptable aqueous carrier and a polysorbate; and (c) instructions for reconstituting the antineoplastic particles into a suspension useful for intracystic injection by: combining the contents of the first vial and the second vial to form the suspension and optionally diluting the suspension with a diluent. In some embodiments the taxane particles have a mean particle size (number) of from 0.1 microns to 1.5 microns. In some embodiments, the taxane particles are paclitaxel particles. In other embodiments, the taxane particles are docetaxel particles.

Antineoplastic Agent Particles

Antineoplastic agents are drugs used to treat neoplasms including malignant pre-cancerous, and non-malignant neoplasms, and include “chemotherapeutic agents”, which are drugs used to treat cancer. Non-limiting examples of antineoplastic agents include taxanes such as paclitaxel, derivatives of paclitaxel, docetaxel, cabazitaxel; epothilones; Vinca alkaloids such as vinblastine, vincristine, vindesine, vinorelbine; camptothecins such as topotecan; platinum complexes such as cisplatin, carboplatin, oxaliplatin; podophyllotoxins such as etoposide and teniposide; and 5-fluorouracil. Other non-limiting examples can be found listed in the “Ashgate Handbook of Antineoplastic Agents”, published by Gower Publishing Limited, 2000, herein incorporated by reference. The antineoplastic agent particles have a mean particle size (number) of from about 0.1 microns to about 5 microns (about 100 nm to about 5000 nm) in diameter. In some embodiments, the antineoplastic agent particles have a mean particle size (number) of from about 0.1 microns to about 1.5 microns (about 100 nm to about 1500 nm) in diameter. In some embodiments, the antineoplastic agent particles have a mean particle size (number) of from about 0.1 microns to less than 1 micron (about 100 nm to less than 1000 nm) in diameter.

In preferred embodiments, the antineoplastic particles are solid, uncoated (“neat” or “naked”) individual particles. In some embodiments, the antineoplastic particles are not bound to any substance. In some embodiments, no substances are absorbed or adsorbed onto the surface of the antineoplastic particles. In some embodiments, the antineoplastic agents or antineoplastic particles are not encapsulated, contained, enclosed or embedded within any substance. In some embodiments, the antineoplastic particles are not coated with any substance. In some embodiments, the antineoplastic particles are not microemulsions, nanoemulsions, microspheres, or liposomes containing an antineoplastic agent. In some embodiments, the antineoplastic particles are not bound to, encapsulated in, or coated with a monomer, a polymer (or biocompatible polymer), a protein, a surfactant, or albumin. In some embodiments, a monomer, a polymer or biocompatible polymer), a protein, a surfactant, or albumin is not absorbed or adsorbed onto the surface of the antineoplastic particles. In some embodiments, the antineoplastic particles are in crystalline form. In other embodiments, the antineoplastic particles are in amorphous form, or a combination of both crystalline and amorphous form. In some embodiments, the antineoplastic particles of the invention contain traces of impurities and byproducts typically found during preparation of the antineoplastic agent. In some embodiments, the antineoplastic particles comprise at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the antineoplastic agent, meaning the antineoplastic particles consist of or consist essentially of substantially pure antineoplastic agent.

In a preferred embodiment, the antineoplastic particles are taxane particles. Taxanes are poorly water soluble compounds generally having a solubility of less than or equal to 10 mg/mL in water at room temperature. Taxanes are widely used as antineoplastic agents and chemotherapy agents. The term “taxanes” as used herein include paclitaxel (I), docetaxel (II), cabazitaxel (III), and any other taxane tar taxane derivatives, non-limiting examples of which are taxol B (cephalomannine), taxol C, taxol D, taxol E, taxol F, taxol G, taxadiene, baccatin III, 10-deacetylbaccatin, taxchinin A, brevifoliol, and taxuspine D, and also include pharmaceutically acceptable salts of taxanes.

Paclitaxel and docetaxel active pharmaceutical ingredients (APIs) are commercially available from Phyton Biotech LLC, Vancouver, Canada. The docetaxel API contains not less than 90%, or not less than 95%, or not less than 97.5% docetaxel calculated on the anhydrous, solvent-free bass. The paclitaxel API contains not less than 90%, or not less than 95%, or not less than 97% paclitaxel calculated on the anhydrous, solvent-free basis. In some embodiments, the paclitaxel API and docetaxel API are USP and/or EP grade. Paclitaxel API can be prepared from a semisynthetic chemical process or from a natural source such as plant cell fermentation or extraction. Paclitaxel is also sometimes referred to by the trade name TAXOL, although this is a misnomer because TAXOL is the trade name of a solution of paclitaxel in polyoxyethylated castor oil and ethanol intended for dilution with a suitable parenteral fluid prior to intravenous infusion. Taxane APIs can be used to make taxane particles. The taxane particles can be paclitaxel particles, docetaxel particles, or cabazitaxel particles, or particles of other taxane derivatives, including particles of pharmaceutically acceptable salts of taxanes.

Taxane particles have a mean particle size (number) of from about 0.1 microns to about 5 microns (about 100 nm to about 5000 nm) in diameter. In some embodiments, the taxane particles have a mean particle size (number) of from about 0.1 microns to about 1.5 microns (about 100 nm to about 1500 nm) in diameter. In some embodiments, the taxane particles have a mean particle size (number) of from about 0.1 microns to less than micron (about 100 nm to less than 1000 nm) in diameter. In preferred embodiments, the taxane particles are solid, uncoated (neat) individual particles. In some embodiments, the taxane particles are not bound to any substance. In some embodiments, no substances are absorbed or adsorbed onto the surface of the taxane particles. In some embodiments, the taxane or taxane particles are not encapsulated, contained, enclosed or embedded within any substance. In some embodiments, the taxane particles are not coated with any substance. In some embodiments, the taxane particles are not microemulsions, nanoemulsions, microspheres, or liposomes containing a taxane. In some embodiments, the taxane particles are not bound to, encapsulated in, or coated with a monomer, a polymer (or biocompatible polymer), a protein, a surfactant, or albumin. In some embodiments, a monomer, a polymer (or biocompatible polymer), a protein, a surfactant, or albumin is not absorbed or adsorbed onto the surface of the taxane particles. In some embodiments, the taxane particles exclude albumin. In some embodiments, the taxane particles are paclitaxel particles and exclude albumin. In some embodiments, the taxane particles are in crystalline form. In other embodiments, the taxane particles are in amorphous form, or a combination of both crystalline and amorphous form. In some embodiments, the taxane particles of the invention contain traces of impurities and byproducts typically found during preparation of the taxane. In some embodiments, the taxane particles comprise at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the taxane, meaning the taxane particles consist of or consist essentially of substantially pure taxane.

The antineoplastic particles or taxane particles (including paclitaxel particles, docetaxel particles, or cabazitaxel particles) can have a mean particle size (number) of from 0.1 microns to 5 microns, or from 0.1 microns to 2 microns, or from 0.1 microns to 1.5 microns, or from 0.1 microns to 1.2 microns, or from 0.1 microns to 1 micron, or from 0.1 microns to less than 1 micron, or from 0.1 microns to 0.9 microns, or from 0.1 microns to 0.8 microns, or from 0.1 to 0.7 microns, or from 0.2 microns to 5 microns, or from 0.2 microns to 2 microns, or from 0.2 microns to 1.5 microns, or from 0.2 microns to 1.2 microns, or from 0.2 microns to 1 micron, or from 0.2 microns to less than 1 micron, or from 0.2 microns to 0.9 microns, or from 0.2 microns to 0.8 microns, or from 0.2 microns to 0.7 microns, or from 0.3 microns to 5 microns, or from 0.3 microns to 2 microns, or from 0.3 microns to 1.5 microns, or from 0.3 microns to 1.2 microns, or from 0.3 microns to 1 micron, or from 0.3 microns to less than 1 micron, or from 0.3 microns to 0.9 microns, or from 0.3 microns to 0.8 microns, or from 0.3 microns to 0.7 microns, or from 0.4 microns to 5 microns, or from 0.4 microns to 2 microns, or from 0.4 microns to 1.5 microns, or from 0.4 microns to 1.2 microns, or from 0.4 microns to 1 micron, or from 0.4 microns to less than 1 micron, or from 0.4 microns to 0.9 microns, or from 0.4 microns to 0.8 microns, or from 0.4 microns to 0.7 microns, or from 0.5 microns to 5 microns, or from 0.5 microns to 2 microns, or from 0.5 microns to 1.5 microns, or from 0.5 microns to 1.2 microns, or from 0.5 microns to 1 micron, or from 0.5 microns to less than 1 micron, or from 0.5 microns to 0.9 microns, or from 0.5 microns to 0.8 microns, or from 0.5 microns to 0.7 microns, or from 0.6 microns to 5 microns, or from 0.6 microns to 2 microns, or from 0.6 microns to 1.5 microns, or from 0.6 microns to 1.2 microns, or from 0.6 microns to 1 micron, or from 0.6 microns to less than 1 micron, or from 0.6 microns to 0.9 microns, or from 0.6 microns to 0.8 microns, or from 0.6 microns to 0.7 microns.

The particle size of the antineoplastic particles including taxane particles can be determined by a particle size analyzer instrument and the measurement is expressed as the mean diameter based on a number distribution (number). A suitable particle size analyzer instrument is one which employs the analytical technique of light obscuration, also referred to as photozone or single particle optical sensing (SPOS). A suitable light obscuration particle size analyzer instrument is the ACCUSIZER, such as the ACCUSIZER 780 SIS, available from Particle Sizing Systems, Port Richey, Florida. Another suitable particle size analyzer instrument is one which employs laser diffraction, such as the Shimadzu SALD-7101.

Antineoplastic agent particles including taxane particles can be manufactured using various particle size-reduction methods and equipment known in the art. Such methods include, but are not limited to conventional particle size-reduction methods such as wet or dry milling, micronizing, disintegrating, and pulverizing. Other methods include “precipitation with compressed anti-solvents” (PCA) such as with supercritical carbon dioxide. In various embodiments, the antineoplastic and/or taxane particles are made by PCA methods as disclosed in U.S. Pat. Nos. 5,874,029, 5,833,891, 6,113,795, 7,744,923, 8,778,181, 9,233,348; US publications US 2015/0375153, US 2016/0354336, US 2016/0374953; and international patent application publications WO 2016/197091, WO 2016/97100, and WO 2016/197101; all of which are herein incorporated by reference.

In PCA particle size reduction methods using supercritical carbon dioxide, supercritical carbon dioxide (anti-solvent) and solvent, e.g. acetone or ethanol, are employed to generate uncoated antineoplastic taxane particles as small as 0.1 to 5 microns within a well-characterized particle-size distribution. The carbon dioxide and solvent are removed during processing (up to 0.5% residual solvent may remain), leaving antineoplastic or taxane particles as a powder. Stability studies show that the paclitaxel particle powder is stable in a vial dose form when stored at room temperature for up to 59 months and under accelerated conditions (40° C./75% relative humidity) for up to six months.

Taxane particles produced by various supercritical carbon dioxide particle size reduction methods can have unique physical characteristics as compared to taxane particles produced by conventional particle size reduction methods using physical impacting or grinding, e.g., wet or dry milling, micronizing, disintegrating, comminuting, microfluidizing, or pulverizing. As disclosed in US publication 2016/0374953, herein incorporated by reference, such unique characteristics include a bulk density (not tapped) between 0.05 g/cm³ and 0.15 g/cm³ and a specific surface area (SSA) of at least 18 m²/g of taxane paclitaxel and docetaxel) particles, which are produced by the supercritical carbon dioxide particle size reduction methods described in US publication 2016/0374953 and as described below. This bulk density range is generally lower than the bulk density of taxane particles produced by conventional means, and the SSA is generally higher than the SSA of taxane particles produced by conventional means. These unique characteristics result in significant increases in dissolution rates in water/methanol media as compared to taxanes produced by conventional means. As used herein, the “specific surface area” (SSA) is the total surface area of the taxane particle per unit of taxane mass as measured by the Brunauer-Emmett-Teller (“BET”) isotherm by the following method: a known mass between 200 and 300 mg of the analyte is added to a 30 mL sample tube. The loaded tube is then mounted to a Porous Materials Inc. SORPTOMETER®, model BET-202A. The automated test is then carried out using the BETWIN® software package and the surface area of each sample is subsequently calculated. As will be understood by those of skill in the art, the “taxane particles” can include both agglomerated taxane particles and non-agglomerated taxane particles; since the SSA is determined on a per gram basis it takes into account both agglomerated and non-agglomerated taxane particles in the composition. The BET specific surface area test procedure is a compendial method included in both the United States Pharmaceopeia and the European Pharmaceopeia. The bulk density measurement can be conducted by pouring the taxane particles into a graduated cylinder without tapping at room temperature, measuring the mass and volume, and calculating the bulk density.

As disclosed in US publication 2016/0374953, studies showed a SSA of 15.0 m²/g and a bulk density of 0.31 g/cm³ for paclitaxel particles produced by milling paclitaxel in a Deco-PBM-V-0.41 ball mill suing a 5 mm ball size, at 600 RPM for 60 minutes at room temperature. Also disclosed in US publication 2016/0374953, one lot of paclitaxel particles had a SSA of 37.7 m²/g and a bulk density of 0.085 g/cm³ when produced by a supercritical carbon dioxide method using the following method: a solution of 65 mg/mL of paclitaxel was prepared in acetone. A BETE MicroWhirl® fog nozzle (BETE Fog Nozzle, Inc.) and a sonic probe (Qsonica, model number Q700) were positioned in the crystallization chamber approximately 8 mm apart. A stainless steel mesh filter with approximately 100 nm holes was attached to the crystallization chamber to collect the precipitated paclitaxel particles. The supercritical carbon dioxide was placed in the crystallization chamber of the manufacturing equipment and brought to approximately 1200 psi at about 38° C. and a flow rate of 24 kg/hour. The sonic probe was adjusted to 60% of total output power at a frequency of 20 kHz. The acetone solution containing the paclitaxel was pumped through the nozzle at a flow rate of 4.5 mL/minute for approximately 36 hours. Additional lots of paclitaxel particles produced by the supercritical carbon dioxide method described above had SSA values of: 22.27 m²/g, 23.90 m²/g, 26.19 m²/g, 30.02 m²/g, 31.16 m²/g, 31.70 m²/g, 32.59 m²/g, 33.82 m²/g, 35.90 m²/g, 38.22 m²/g, and 38.52 m²/g.

As disclosed in US publication 201610374953, studies showed a SSA of 15.2 m²/g and a bulk density of 0.44 g/cm³ for docetaxel particles produced by milling docetaxel in a Deco-PBM-V-0.41 ball mill suing a 5 mm ball size, at 600 RPM for 60 minutes at room temperature. Also disclosed in US publication 2016/0374953, docetaxel particles had a SSA of 44.2 m²/g and a bulk density of 0.079 g/cm³ when produced by a supercritical carbon dioxide method using the following method: A solution of 79.32 mg/mL of docetaxel was prepared in ethanol. The nozzle and a sonic probe were positioned in the pressurizable chamber approximately 9 mm apart. A stainless steel mesh filter with approximately 100 nm holes was attached to the pressurizable chamber to collect the precipitated docetaxel particles. The supercritical carbon dioxide was placed in the pressurizable chamber of the manufacturing equipment and brought to approximately 1200 psi at about 38° C. and a flow rate of 68 slpm. The sonic probe was adjusted to 60% of total output power at a frequency of 20 kHz. The ethanol solution containing the docetaxel was pumped through the nozzle at a flow rate of 2 mL/minute for approximately 95 minutes). The precipitated docetaxel agglomerates and particles were then collected from the supercritical carbon dioxide as the mixture is pumped through the stainless steel mesh filter. The filter containing the particles of docetaxel was opened and the resulting product was collected from the filter.

As disclosed in US publication 2016/0374953, dissolution studies showed an increased dissolution rate in methanol/water media of paclitaxel and docetaxel particles made by the supercritical carbon dioxide methods described in US publication 2016/0374953 as compared to paclitaxel and docetaxel particles made by minima paclitaxel and docetaxel using a Deco-PBM-V-0.41 ball mill suing a 5 mm ball size, at 600 RPM for 60 minutes at room temperature. The procedures used to determine the dissolution rates are as follows. For paclitaxel, approximately 50 mg of material were coated on approximately 1.5 grams of 1 mm glass beads by tumbling the material and beads in a vial for approximately 1 hour. Beads were transferred to a stainless steel mesh container and placed in the dissolution bath containing methanol/water 50/50 (v/v) media at 37° C., pH 7, and a USP Apparatus II (Paddle), operating at 75 rpm. At 10, 20, 30, 60, and 90 minutes, a 5 mL aliquot was removed, filtered through a 0.22 μm filter and analyzed on a UV/VIS spectrophotometer at 227 nm. Absorbance values of the samples were compared to those of standard solutions prepared in dissolution media to determine the amount of material dissolved. For docetaxel, approximately 50 mg of material was placed directly in the dissolution bath containing methanol/water 15/85 (v/v) media at 37° C., pH 7, and a USP Apparatus II (Paddle), operating at 75 rpm. At 5, 15, 30, 60, 120 and 225 minutes, a 5 mL aliquot was removed, filtered through a 0.22 μm filter, and analyzed on a UV/VIS spectrophotometer at 232 nm. Absorbance values of the samples were compared to those of standard solutions prepared in dissolution media to determine the amount of material dissolved. For paclitaxel, the dissolution rate was 47% dissolved in 30 minutes for the particles made by the supercritical carbon dioxide method versus 32% dissolved in 30 minutes for the particles made by milling. For docetaxel, the dissolution rate was 27% dissolved in 30 minutes for the particles made by the supercritical carbon dioxide method versus 9% dissolved in 30 minutes for the particles made by milling.

In some embodiments, the antineoplastic particles have a SSA of at least 10, at least 12, at least 14, at least 16, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, or at least 35 m²/g. In one embodiment, the antineoplastic particles have an SSA of between about 10 m²/g and about 50 m²/g. In some embodiments, the antineoplastic particles have a bulk density between about 0.050 g/cm³ and about 0.20 g/cm³.

In further embodiments, the antineoplastic particles have a SSA of:

-   -   (a) between 16 m²/g and 31 m²/g or between 32 m²/g and 40 m²/g;     -   (b) between 16 m²/g and 30 m²/g or between 32 m²/g and 40 m²/g;     -   (c) between 16 m²/g and 29 m²/g or between 32 m²/g and 40 m²/g;     -   (d) between 17 m²/g and 31 m²/g or between 32 m²/g and 40 m²/g;     -   (e) between 17 m²/g and 30 m²/g or between 32 m²/g and 40 m²/g;     -   (f) between 17 m²/g and 29 m²/g or between 32 m²/g and 40 m²/g;     -   (g) between 16 m²/g and 31 m²/g or between 33 m²/g and 40 m²/g;     -   (h) between 16 m²/g and 30 m²/g or between 33 m²/g and 40 m²/g;     -   (i) between 16 m²/g and 29 m²/g or between 33 m²/g and 40 m²/g;     -   (j) between 17 m²/g and 31 m²/g or between 33 m²/g and 40 m²/g;     -   (k) between 17 m²/g and 30 m²/g or between 33 m²/g and 40 m²/g;     -   (l) between 17 m²/g and 29 m²/g, or between 33 m²/g and 40 m²/g;     -   (m) between 16 m²/g and 31 m²/g, or ≥32 m²/g;     -   (h) between 17 m²/g and 31 m²/g, or ≥32 m²/g;     -   (i) between 16 m²/g and 30 m²/g, or ≥32 m²/g;     -   (j) between 17 m²/g and 30 m²/g, or ≥32 m²/g;     -   (k) between 16 m²/g and 29 m²/g, or ≥32 m²/g;     -   (l) between 17 m²/g and 29 m²/g, or ≥32 m²/g;     -   (m) between 16 m²/g and 31 m²/g, or ≥33 m²/g;     -   (n) between 17 m²/g and 31 m²/g, or ≥33 m²/g;     -   (o) between 16 m²/g and 30 m²/g, or ≥33 m²/g;     -   (p) between 17 m²/g and 30 m²/g, or ≥33 m²/g;     -   (q) between 16 m²/g and 29 m²/g, or ≥33 m²/g; or     -   (r) between 17 m²/g and 29 m²/g, or ≥33 m²/g.

In some embodiments, the antineoplastic particles are taxane particles. In some embodiments, the antineoplastic particles or taxane particles are non-agglomerated individual particles and are not clusters of multiple antineoplastic or taxane particles.

In some embodiments, the taxane particles are paclitaxel particles and have an SSA of at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, or at least 35 m²/g. In other embodiments, the paclitaxel particles have an SSA of 18 m²/g to 50 m²/g, or 20 m²/g, to 50 m²/g, or 22 m²/g to 50 m²/g or 25 m²/g to 50 m²/g, or 26 m²/g to 50 m²/g, or 30 m²/g to 50 m²/g, or 35 m²/g to 50 m²/g, or 18 m²/g to 45 m²/g, or 20 m²/g to 45 m²/g, or 22 m²/g to 45 m²/g, or 25 m²/g to 45 m²/g, or 26 m²/g to 45 m²/g or 30 m²/g to 45 m²/g, or 35 m²/g to 45 m²/g, or 18 m²/g to 40 m²/g, or 20 m²/g to 40 m²/g, or 22 m²/g to 40 m²/g, or 25 m²/g to 40 m²/g, or 26 m²/g to 40 m²/g, or 30 m²/g to 40 m²/g, or 35 m²/g to 40 m²/g.

In some embodiments, the paclitaxel particles have a bulk density (not-tapped) of 0.05 g/cm³ to 0.15 g/cm³, or 0.05 g/cm³ to 0.20 g/cm³.

In some embodiments, the paclitaxel particles have a dissolution rate of at least 40% w/w dissolved in 30 minutes or less in a solution of 50% methanol/50% water (v/v) in a USP II paddle apparatus operating at 75 RPM, at 37° C., and at a pH of 7.

In some embodiments, the taxane particles are docetaxel particles and have an SSA of at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, at least 40, at least 41, or at least 42 m²/g. In other embodiments, the docetaxel particles have an SSA of 18 m²/g to 60 m²/g, or 22 m²/g to 60 m²/g, or 25 m²/g to 60 m²/g, or 30 m²/g to 60 m²/g, or 40 m²/g to 60 m²/g, or 18 m²/g to 50 m²/g, or 22 m²/g to 50 m²/g, or 25 m²/g to 50 m²/g, or 26 m²/g to 50 m²/g, or 30 m²/g to 50 m²/g, or 35 m²/g to 50 m²/g, or 40 m²/g to 50 m²/g.

In some embodiments, the docetaxel particles have a bulk density (not-tapped) of 0.05 g/cm³ to 0.15 g/cm³.

In some embodiments, the docetaxel particles have a dissolution rate of at least 20% w/w dissolved in 30 minutes or less in a solution of 15% methanol/85% water (v/v) in a USP II paddle apparatus operating at 75 RPM, at 37° C., and at a pH of 7.

The antineoplastic particles, including taxane particles, can be packaged into any suitable container such as glass or plastic vials. A non-limiting example of a suitable container is a Type 1, USP, clear-glass vial closed with a bromobutyl rubber stopper and aluminum crimp seal. The antineoplastic particles can be sterilized after the particles are the container using sterilization methods known in the art such as gamma irradiation or autoclaving.

Compositions

The compositions of the invention comprise antineoplastic particles, such as taxane particles. The compositions can further comprise a carrier. The carrier can be a liquid (fluid) carrier, such as an aqueous carrier. Non-limiting examples of suitable aqueous carriers include water, such as Sterile Water for Injection USP; 0.9% saline solution (normal saline), such as 0.9% Sodium Chloride for Injection USP; dextrose solution, such as 5% Dextrose for Injection USP; and Lactated Ringer's Solution for Injection USP. Non-aqueous based liquid carriers and other aqueous-based liquid carriers can be used. The carrier can be a pharmaceutically acceptable carrier, i.e., suitable for administration to a subject by injection or other routes of administration. The carrier can be any other type of liquid such as emulsions or flowable semi-solids. Non-limiting examples of flowable semisolids include gels and thermosetting gels. The composition can be a suspension, i.e., a suspension dosage form composition where the antineoplastic particles, such as taxane particles, are dispersed (suspended) within a continuous carrier/and or diluent. The antineoplastic particles can be completely dispersed, partially dispersed and partially dissolved, but not completely dissolved in the carrier. In some embodiments, the composition is a suspension of taxane particles dispersed within a continuous carrier. In a preferred embodiment, the carrier is a pharmaceutically acceptable carrier. In preferred embodiments, the composition is sterile. In various embodiments, the composition comprises, consists essentially of, or consists of antineoplastic particles and a liquid carrier, wherein the composition is a suspension of the antineoplastic particles dispersed within the liquid carrier. In some embodiments, the composition consists essentially of or consists of antineoplastic particles and a carrier, wherein the carrier is an aqueous carrier and wherein the composition is a suspension.

The composition of antineoplastic particles and a carrier can be administered as-is. Optionally, the composition of antineoplastic particles and a carrier can further comprise a suitable diluent to dilute the composition in order to achieve a desired concentration (dose) of antineoplastic particles. In some embodiments, the carrier can serve as the diluent; stated another way, the amount of carrier in the composition provides the desired concentration of antineoplastic particles in the composition and no further dilution is needed. A suitable diluent can be a fluid, such as an aqueous fluid. Non-limiting examples of suitable aqueous diluents include water, such as Sterile Water for Injection USP; 0.9% saline solution (normal saline), such as 0.9% Sodium Chloride for Injection USP; dextrose solution, such as 5% Dextrose for Injection USP; and Lactated Ringer's Solution for Injection USP. Other liquid and aqueous-based diluents suitable for administration by injection can be used and can optionally include salts, buffering agents, and/or other excipients. In some embodiments, the diluent is sterile. The composition can be diluted with the diluent at a ratio to provide a desired concentration dosage of the antineoplastic particles. For example, the volume ratio of composition to diluent might be in the range of 1:1-1:100 v/v or other suitable ratios. In some embodiments, the composition comprises antineoplastic particles, a carrier, and a diluent, wherein the carrier and diluent form a mixture, and wherein the composition is a suspension of antineoplastic particles dispersed in the carrier/diluent mixture. In some embodiments, the carrier/diluent mixture is a continuous phase and the antineoplastic particles are a dispersed phase.

The composition, carrier, and/or diluent can further comprise functional ingredients such as buffers, salts, osmotic agents, surfactants, viscosity modifiers, rheology modifiers, suspending agents, pH adjusting agents such as alkalinizing agents or acidifying agents, tonicity adjusting agents, preservatives, antimicrobial agents including quaternary ammonium compounds such as benzalkonium chloride and benzethonium chloride, demulcents, antioxidants, antifoaming agents, alcohols such as ethanol, chelating agents, and/or colorants. For example, the composition can comprise taxane particles and a carrier comprising water, a salt, a surfactant, and optionally a buffer. In one embodiment, the carrier is an aqueous carrier and comprises a surfactant, wherein the concentration of the surfactant is 1% or less on a w/w or w/v basis; in other embodiments, the surfactant is less than 0.5%, less than 0.25%, less than 0.1%, or about 0.1%. In other embodiments, the aqueous carrier excludes the surfactants GELUCIRE® (polyethylene glycol glycerides composed of mono-, di- and triglycerides and mono- and diesters of polyethylene glycol) and/or CREMOPHOR® (polyethoxylated castor oil). In some embodiments, the composition or carrier excludes polymers, proteins (such as albumin), polyethoxylated castor oil, and/or polyethylene glycol glycerides composed of mono-, di- and triglycerides and mono- and diesters of polyethylene glycol.

The composition, carrier, and/or diluent can comprise one or more surfactants. Suitable surfactants include by way of example and without limitation polysorbates, lauryl sulfates, acetylated monoglycerides, diacetylated monoglycerides, and poloxamers, such as poloxamer 407. Polysorbates are polyoxyethylene sorbitan fatty acid esters which are a series of partial fatty acid esters of sorbitol and its anhydrides copolymerized with approximately 20, 5, or 4 moles of ethylene oxide for each mole of sorbitol and its anhydrides. Non-limiting examples of polysorbates are polysorbate 20, polysorbate 21, polysorbate 40, polysorbate 60, polysorbate 61, polysorbate 65, polysorbate 80, polysorbate 81, polysorbate 85, and polysorbate 120. Polysorbates containing approximately 20 moles of ethylene oxide are hydrophilic nonionic surfactants. Examples of polysorbates containing approximately 20 moles of ethylene oxide include polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, polysorbate 85, and polysorbate 120. Polysorbates are available commercially from Croda under the tradename TWEEN™. The number designation of the polysorbate corresponds to the number designation of the TWEEN, e.g., polysorbate 20 is TWEEN 20, polysorbate 40 is TWEEN 40, polysorbate 60 is TWEEN 60, polysorbate 80 is TWEEN 80, etc. USP/NF grades of polysorbate include polysorbate 20 NF, polysorbate 40 NF, polysorbate 60 NF, and polysorbate 80 NF. Polysorbates are also available in PhEur grades (European Pharmacopoeia), BP grades, and JP grades. The term “polysorbate” is a non-proprietary name. The chemical name of polysorbate 20 is polyoxyethylene 20 sorbitan monolaurate. The chemical name of polysorbate 40 is polyoxyethylene 20 sorbitan monopalmitate. The chemical name of polysorbate 60 is polyoxyethylene 20 sorbitan monostearate. The chemical name of polysorbate 80 is polyoxyethylene 20 sorbitan monooleate. In some embodiments, the composition, carrier, and/or diluent can comprise mixtures of polysorbates. In some embodiments, the composition, carrier, and/or diluent comprises polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, polysorbate 85, and/or polysorbate 120. In some embodiments, the composition, carrier, and/or diluent comprises polysorbate 20, polysorbate 40, polysorbate 60, and/or polysorbate 80. In one embodiment, the composition, carrier, and/or diluent comprises polysorbate 80.

In some embodiments, the composition, carrier, and/or diluent can comprise an alcohol, such as ethanol. The ethanol can be USP grade such as Alcohol USP or Dehydrated Alcohol (200 proof) USP. In some embodiments, the composition comprises taxane particles, a carrier, and optionally a diluent, wherein the carrier and/or diluent comprises water, ethanol, and a polysorbate. In some embodiments, the ethanol is present in the composition, carrier, and/or diluent at a concentration of about 0.1% w/v to about 10% w/v, or about 0.1% w/v to about 8% w/v, or about 2% w/v to about 8% w/v, or about 5% w/v to about 10% w/v, or about 8% w/v. In some embodiments, the ethanol is present in the composition at a concentration of about 0.1 w/v to about 4% w/v, or about 2% w/v to about 4% w/v, or about 3.2% w/v.

In some embodiments, the composition comprises antineoplastic particles, a carrier, and optionally a diluent, wherein the carrier an for diluent comprises water and a polysorbate. In one embodiment, the composition is a suspension, the antineoplastic particles are taxane particles, and the polysorbate is polysorbate 80. In other embodiments, the polysorbate or polysorbate 80 is present in the composition, carrier, and/or diluent at a concentration of between about 0.01% v/v and about 1.5% v/v. The inventors have surprisingly discovered that the recited very small amounts of polysorbate 80 reduce the surface tension at the interface of the antineoplastic particles and the aqueous carrier (such as saline solution). These embodiments are typically formulated near the time of use of the composition. In some embodiments, the particles may be coated with the polysorbate or polysorbate 80. In other embodiments, the particles are not coated with the polysorbate. polysorbate 80. In various other embodiments, the polysorbate or polysorbate 80 is present in the composition, carrier, and/or diluent at a concentration of between: about 0.01% v/v and about 1% v/v, about 0.01% v/v and about 0.5% v/v, about 0.01% v/v and about 0.4% v/v, about 0.01% v/v and about 0.35% v/v, about 0.01% v/v and about 0.3% v/v, about 0.01% v/v and about 0.25% v/v, about 0.01% v/v and about 0.2% v/v, about 0.01% v/v and about 0.15% v/v, about 0.01% v/v and about 0.1% v/v, about 0.05% v/v and about 1% v/v, about 0.05% v/v and about 0.5% v/v, about 0.05% v/v and about 0.4% v/v, about 0.05% v/v and about 0.35% v/v, about 0.05% v/v and about 0.3% v/v, about 0.05% v/v and about 0.25% v/v, about 0.05% v/v and about 0.2% v/v, about 0.05% v/v and about 0.15% v/v, about 0.05% v/v and about 0.1% v/v, about 0.1% v/v and about 1% v/v, about 0.1% v/v and about 0.5% v/v, about 0.1% v/v and about 0.4% v/v, about 0.1% v/v and about 0.35% v/v, about 0.1% v/v and about 0.3% v/v, about 0.1% v/v and about 0.25% v/v, about 0.1% v/v and about 0.2% v/v, about 0.1% v/v and about 0.15% v/v, about 0.2% v/v and about 1% v/v, about 0.2% v/v and about 0.5% v/v, about 0.2% v/v and about 0.4% v/v, about 0.2% v/v and about 0.35% v/v, about 0.2% v/v and about 0.3% v/v, about 0.2% v/v and about 0.25% v/v, about 0.3% v/v and about 1% v/v, about 0.3% v/v and about 0.5% v/v, about 0.3% v/v and about 0.4% v/v, or about 0.3% v/v and about 0.35% v/v; or about 0.01%, about 0.05%, about 0.1% v/v, about 0.15% v/v, about 0.16% v/v, about 0.2% v/v, about 0.25% v/v, about 0.3% v/v, about 0.35% v/v, about 0.4% v/v, about 0.45% v/v, about 0.5% v/v, or about 1% v/v.

The composition, carrier, and/or diluent can comprise one or more tonicity adjusting agents. Suitable tonicity adjusting agents include by way of example and without limitation, one or more inorganic salts, electrolytes, sodium chloride, potassium chloride, sodium phosphate, potassium phosphate, sodium, potassium sulfates, sodium and potassium bicarbonates and alkaline earth metal salts, such as alkaline earth metal inorganic salts, e.g., calcium salts, and magnesium salts, mannitol, dextrose, glycerin, propylene glycol, and mixtures thereof.

The composition, carrier, and/or diluent can comprise one or more buffering agents. Suitable buffering agents include by way of example and without limitation, dibasic sodium phosphate, monobasic sodium phosphate, citric acid, sodium citrate, tris(hydroxymethyl)aminomethane, bis(2-hydroxyethyl)iminotris-(hydroxymethyl)methane, and sodium hydrogen carbonate and others known to those of ordinary skill in the art. Buffers are commonly used to adjust the pH to a desirable range for intraperitoneal use. Usually a pH of around 5 to 9, 5 to 8, 6 to 7.4, 6.5 to 7.5, or 6.9 to 7.4 is desired.

The composition, carrier, and/or diluent can comprise one or more demulcents. A demulcent is an agent that forms a soothing film over a mucous membrane, such as the membranes lining the peritoneum and organs therein. A demulcent may relieve minor pain and inflammation and is sometimes referred to mucoprotective agent. Suitable demulcents include cellulose derivatives ranging from about 0.2 to about 2.5% such as carboxymethylcellulose sodium, hydroxyethyl cellulose, hydroxypropyl methylcellulose, and methylcellulose; gelatin at about 0.01%; polyols in about 0.05 to about 1%, also including about 0.05 to about 1%, such as glycerin, polyethylene glycol 300, polyethylene glycol 400, and propylene glycol; polyvinyl alcohol from about 0.1 to about 4%; povidone from about 0.1 to about 2%; and dextran 70 from about 0.1% when used with another polymeric demulcent described herein.

The composition, carrier, and/or diluent can comprise one or more alkalinizing agents to adjust the pH. As used herein, the term “alkalizing agent” is intended to mean a compound used to provide an alkaline medium. Such compounds include, by way of example and without limitation, ammonia solution, ammonium carbonate, potassium hydroxide, sodium carbonate, sodium bicarbonate, and sodium hydroxide and others known to those of ordinary skill in the art.

The composition, carrier, and/or diluent can comprise acidifying agents to adjust the pH. As used herein, the term “acidifying agent” is intended to mean a compound used to provide an acidic medium. Such compounds include, by way of example and without limitation, acetic acid, amino acid, citric acid, nitric acid, fumaric acid and other alpha hydroxy acids, hydrochloric acid, ascorbic acid, and nitric acid and others known to those of ordinary skill in the art.

The composition, carrier, and/or diluent can comprise one or more antifoaming agents. As used herein, the term “antifoaming agent”, intended to mean a compound or compounds that prevents or reduces the amount of foaming that forms on the surface of the fill composition. Suitable antifoaming agents include by way of example and without limitation, dimethicone, SIMETHICONE, octoxynol and others known to those of ordinary skill in the art.

The composition, carrier, and/or diluent can comprise one or more viscosity modifier that increase or decrease the viscosity of the suspension. Suitable viscosity modifiers include methylcellulose, hydroxypropyl methycellulose, mannitol, polyyinylpyrrolidone, cross-linked acrylic acid polymers such as carbomer, and others known to those of ordinary skill in the art. The composition, carrier, and/or diluent can further comprise rheology modifiers to modify the flow characteristics of the composition to allow it to adequately flow through devices such as injection needles or tubes. Non-limiting examples of viscosity and rheology modifiers can be found in “Rheology Modifiers Handbook—Practical Use and Application” Braun, William Andrew Publishing, 2000.

The concentration or amount of antineoplastic particles in the composition or dosage is at an “effective amount”, i.e., to provide a therapeutic effect on an epithelial cyst of a subject by accomplishing one or more of the following: (a) reducing cyst volume/size; (b) reducing cyst growth rate; (c) eliminating a cyst; (d) ablation of a cyst or (e) reducing pain associated with the cyst. In one embodiment, the concentration of the antineoplastic particles, which can be taxane particles, in the composition is between about 0.1 mg/mL and about 100 mg/mL. In various further embodiments, the concentration of antineoplastic particles, which can be taxane particles, in the composition is between: about 0.5 mg/mL and about 100 mg/mL, about 1 mg/mL and about 100 mg/mL, about 2 mg/mL and about 100 mg/mL, about 5 mg/mL and about 100 mg/mL, about 10 mg/mL and about 100 mg/mL, about 25 mg/mL and about 100 mg/mL, about 30 mg/mL and about 100 mg/mL, about 0.1 mg/mL and about 75 mg/mL, about 0.5 mg/mL and about 75 mg/mL, about 1 mg/mL and about 75 mg/mL, about 2 mg/mL and about 75 mg/mL, about 5 mg/mL and about 75 mg/mL, about 10 and about 75 mg/mL, about 25 mg/mL and about 75 mg/mL, about 30 mg/mL and about 75 mg/mL, about 0.1 mg/mL and about 50 mg/mL, about 0.5 mg/mL and about 50 mg/mL, about 1 mg/mL and about 50 mg/mL, about 2 mg/mL and about 50 mg/mL, about 5 mg/mL and about 50 mg/mL, about 10 mg/mL and about 50 mg/mL, about 25 mg/mL and about 50 mg/mL, about 30 mg/mL and about 50 mg/mL, about 0.1 mg/mL and about 40 mg/mL, about 0.5 mg/mL and about 40 mg/mL, about 1 mg/mL and about 40 mg/mL, about 2 mg/mL and about 40 mg/mL, about 5 mg/mL and about 40 mg/mL, about 10 mg/mL and about 40 mg/mL, about 25 mg/mL and about 40 mg/mL, about 30 mg/mL and about 40 mg/mL, about 0.1 mg/mL and about 30 mg/mL, about 0.5 mg/mL and about 30 mg/mL, about 1mg/mL and about 30 mg/mL, about 2 mg/mL and about 30 mg/mL, about 5 mg/mL and about 30 mg/mL, about 10 mg/mL and about 30 mg/mL, about 25 mg/mL and about 30 mg/mL, about 0.1 mg/mL and about 25 mg/mL, about 0.5 mg/mL and about 25 mg/mL, about 1 mg/mL and about 25 mg/mL, about 2 mg/mL and about 25 mg/mL, about 5 mg/mL and about 25 mg/mL, about 10 mg/mL and about 25 mg/mL, about 0.1 mg/mL and about 20 mg/mL, about 0.5 mg/mL and about 20 mg/mL, about 1 mg/mL and about 20 mg/mL, about 2 mg/mL and about 20 mg/mL, about 5 mg/mL and about 20 mg/mL, about 10 mg/mL and about 20 mg/mL, about 0.1 mg/mL and about 15 mg/mL, about 0.5 mg/mL and about 15 mg/mL, about 1 mg/mL and about 15 mg/mL, about 2 mg/mL and about 15 mg/mL, about 5 mg/mL and about 15 mg/mL, about 10 mg/mL and about 15 mg/mL, about 0.1 mg/mL and about 10 mg/mL, about 0.5 mg/mL and about 10 mg/mL, about 1 mg/mL and about 10 mg/mL, about 2 mg/mL and about 10 mg/mL, about 5 mg/mL and about 10 mg/mL, about 0.1 mg/mL and about 5 mg/mL, about 0.5 mg/mL and about 5 mg/mL, about 1 mg/mL and about 5 mg/mL, about 2 mg/mL and about 5 mg/mL, about 0.1 mg/mL and about 2 mg/mL, about 0.5 mg/mL and about 2 mg/mL, about 1 mg/mL and about 2 mg/mL, about 0.1 mg/mL and about 1 mg/mL, about 0.5 mg/mL and about 1 mg/mL, about 0.1 mg/mL and about 0.5 mg/mL, about 3 mg/mL and about 8 mg/mL, or about 4 mg/mL and about 6 mg/mL; or at least about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 61, 65, 70, 75, or 100 mg/mL; or about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 55, 60, 61, 65, 70, 75, or 100 mg/mL. The antineoplastic particles may be the sole therapeutic agent administered, or may be administered with other therapeutic agents.

In various embodiments, the composition comprises taxane particles (paclitaxel particles or docetaxel particles), a carrier, and a diluent, wherein the concentration of taxane particles in the composition (including the carrier and diluent) is between: about 1 mg/mL and about 40 mg/mL, about 5 mg/mL and about 20 mg/mL, about 5 mg/mL and about 15 mg/mL, about 5 mg/mL and about 10 mg/mL, about 6 mg/mL and about 20 mg/mL, about 6 mg/mL and about 15 mg/mL, about 6 mg/mL and about 10 mg/mL, about 10 mg/mL and about 20 mg/mL, or about 10 mg/mL and about 15 mg/mL; or about 6 mg/mL, about 10 mg/mL, or about 15 mg/mL. In further embodiments, the carrier is an aqueous carrier which can be saline solution, such as about 0.9% sodium chloride solution and the diluent is an aqueous diluent which can be saline solution, such as about 0.9% sodium chloride solution. In further embodiments, the aqueous carrier comprises a polysorbate, such as polysorbate 80.

The compositions should be at suitable volume to supply a sufficient dose volume for intracystic injections, i.e., where the volume of the dose is equal to the amount of cyst fluid withdrawn from the cyst prior to injection. Thus, the volume of the composition should be equal to or greater than the dose volume. The volume of withdrawn cyst fluid can vary depending on the type of cyst. Generally, the volume of cyst fluid removed from pancreatic cysts is about 2 to about 10 mL, and typically is about 4 to about 5 mL. In some embodiments, the volume of the composition is sufficient to provide a dose volume of about 2 to about 10 mL, or about 4 to about 5 mL.

In some embodiments, the volume of the composition is: 10 ul-60 mL, 10 ul-50 mL, 10 ul-40 mL, 10 ul-35 mL, 10 ul-30 mL, 10 ul-25 mL, 10 ul-20 mL, 10 ul-15 mL, 10 ul-10 mL, 10 ul-7.5 mL, 10 ul-7 mL, 10 ul-5 mL, 10 ul-4 mL, 10 ul-3 mL, 50 ul-60 mL, 50ul-50 mL, 50 ul-40 mL, 50 ul-35 mL, 50 ul-30 mL, 50 ul-25 mL, 50 ul-20 mL, 50 ul-15 mL, 50 ul-10 mL, 50 ul-7.5 mL 50 ul-7 mL, 50 ul-5 mL, 50 ul-4 mL, 50 ul-3 mL, 100 ul-60 mL, 100 ul-50 mL 100 ul-40 mL, 100 ul-35 mL, 100 ul-30 mL, 100 ul-25 mL, 100 ul-20 mL, 100 ul-15 mL, 100 ul-10 mL, 100 ul-7.5 mL, 100 ul-7 mL, 100 ul-5 mL, 100 ul-4 mL, 100 ul-3 mL, 500 ul-60 mL, 500 ul-50 mL 500 ul-40 mL, 500 ul-35 mL, 500 ul-30 mL, 500 ul-25 mL, 500 ul-20 mL, 500 ul-15 mL, 500 ul-10 mL, 500 ul-7.5 mL, 500 ul-7 mL, 500 ul-5 mL, 500 ul-4 mL, 500 ul-3 mL, 1 mL-60 mL, 1 mL-50 mL 1 mL-40 mL, 1 mL-35 mL, 1 mL-30 mL, 1 mL-25 mL, 1 mL-20 mL, 1 mL-15 mL, 1 mL-12 mL, 1 mL-10 mL, 1 mL-7.5 mL, 1 mL-7 mL, 1 mL-5 mL, 1 mL-4 mL, or 1 mL-3 mL. In some embodiments, the volume of the composition is about: 1 mL, 2 mL, 3 mL, 4, mL, 5 mL, 6, mL, 7 mL, 8, mL, 9 mL, 10 mL, 12 mL, 15, mL, 20 mL, 30 mL, 40, mL, 50 mL, or 60 mL. Thus, in another aspect, the invention provides pharmaceutical compositions comprising antineoplastic particles or taxane particles and a pharmaceutically acceptable carrier, where the total volume of the composition is between: 1 mL and 60 mL, 1 mL and 50 mL, 1 mL and 40 mL, 1 mL and 35 mL, 1 mL and 30 mL, and 1 mL and 25 mL, 1 mL and 20 mL, 1 mL and 15 mL, 1 mL and 12 mL, 1 mL and 10 mL, 1 mL and 7.5 mL, 1 mL and 7 mL, 1 mL and 5 mL, 1 mL and 4 mL, or 1 mL and 3 mL.

Kits

The present invention also provides kits, comprising:

-   -   (a) a first vial comprising, consisting essentially of, or         consisting of antineoplastic particles;     -   (b) a second vial comprising a pharmaceutically acceptable         carrier; and     -   (c) instructions for reconstituting the antineoplastic particles         into a suspension useful for intracystic injection by: combining         the contents of the first vial and the second vial to form the         suspension and optionally diluting the suspension with a         diluent.

In preferred embodiments, the antineoplastic particles are taxane particles such as paclitaxel particles or docetaxel particles. The antineoplastic particles in the first vial can be in a powder form. The antineoplastic particles in the first vial can be the sole ingredient in the first vial. In some embodiments, the taxane particles have a mean particle size (number) of from 0.1 microns to 1.5 microns. The pharmaceutically acceptable carrier can be an aqueous carrier such as 0.9% saline solution. The carrier can further comprise a surfactant such as a polysorbate. In some embodiments, the polysorbate is polysorbate 80. In some embodiments, the polysorbate or polysorbate 80 is at a concentration of between about 0.01% v/v and about 1% v/v. In some embodiments, the taxane particles are paclitaxel particles and have a specific surface area (SSA) of at least 18 m²/g and/or a bulk density (not-tapped) of 0.05 g/cm³ to 0.15 g/cm³. In other embodiments, the taxane particles are docetaxel particles and have a specific surface area (SSA) of at least 18 m²/g and/or a bulk density (not-tapped) of 0.05 g/cm³ to 0.15 g/cm³. When the suspension of antineoplastic particles and carrier containing a polysorbate is diluted with the diluent, excessive dissolution of the antineoplastic particles is prevented.

Any suitable vial can be used in the kits. A non-limiting example of a suitable vial is a Type 1, USP, clear-glass vial closed with a bromobutyl rubber stopper and aluminum crimp seal. The volumes of the vials can vary depending on the amount of antineoplastic particles, the volume of the carrier, and the volume of the final reconstituted suspension. The vials and their contents can be sterilized using sterilization methods known in the art such as gamma irradiation or autoclaving. In some embodiments, the contents of the vials are sterile. The kits can be configured for single-dose or multiple-dose administration.

A non-limiting exemplary procedure for preparing a suspension composition from a kit is as follows:

-   -   1. Using a syringe with a suitable gauge needle, add all or a         portion of the carrier from the second vial into the first vial         containing the anti-neoplastic particles.     -   2. Vigorously hand shake the first vial with inversions to make         sure all the particles adhering to the interior of the vial and         stopper are wetted.     -   3. Continue shaking for 1 minute and examine the suspension for         any large clumps of particles.     -   4. Immediately after shaking, use a syringe with a suitable         gauge needle to add a prescribed volume of a diluent to the         first vial to dilute the suspension to a desired dose level, and         hand shake the vial for another 1 minute. Periodically examine         the suspension for any large visible clumps. If present,         continue hand mixing until the suspension is properly dispersed.     -   5. After mixing, allow the suspension to sit undisturbed for at         least 5 minutes to reduce entrapped air and foam.

The compositions, suspensions, and kits of the invention can include any embodiment or combination of embodiments described herein including any embodiments of the antineoplastic particles, any embodiments of the carriers and diluents, and any embodiments of the polysorbate or polysorbate 80 concentrations. The compositions, suspensions, and kits can exclude polymers, proteins (such as albumin), polyethoxylated castor oil, and/or polyethylene glycol glycerides composed of mono-, di- and triglycerides and mono- and diesters of polyethylene glycol. The compositions and kits can further comprise other components as appropriate for given antineoplastic particles.

Methods of Administration/Treatment

The compositions comprising antineoplastic particles, including taxane particles, described and disclosed supra can be used in methods for the treatment of epithelial cystic neoplasms (cysts) in subjects by direct injection of the compositions into the cysts (intracystic injection). Epithelial cysts are cysts which have an epithelial inner lining. The compositions and methods disclosed herein are useful for treatment of any type of epithelial cyst. Non-limiting examples of epithelial cysts include gastrointestinal cysts such as hepatic cysts, pancreatic cysts, splenic cysts, colon cysts; urologic cysts such as renal cysts, epididymal cysts, prostatic cysts; gynecological cysts such as ovarian cysts and vaginal cysts; head and neck cysts such as thyroid cysts, parathyroid cysts, and other head and neck cysts; as well as other cysts such as Baker's cysts, lung cysts, lymphatic cysts, and pericardial cysts. In some embodiments, the epithelial cyst is a pancreatic cyst. A pancreatic cyst can be an intraductal papillary mucinous neoplasm (IPMN), a mucinous cystic neoplasms (MCN), or a serous cystadenoma. In some embodiments, the pancreatic cyst is an intraductal papillary mucinous neoplasm (IPMN). In other embodiments, the pancreatic cyst is a mucinous cystic neoplasms (MCN). In still other embodiments, the pancreatic cyst is a serous cystadenoma.

The injection of the composition into an epithelial cyst can be conducted by use of a procedure known as “endoscopic ultrasound-guided fine needle injection” (EUS-FNI), which is a procedure in which endoscopy is combined with ultrasound to aid in the location of the cyst and to facilitate the injection of the composition into the cyst. Because the antineoplastic particles are solid particles, the suspensions of antineoplastic particles can be more easily visualized with sonography than solutions of antineoplastic agents or even suspensions of albumin coated particles. This is especially evident when the antineoplastic particles are in crystalline form. This feature provides a tremendous advantage in that the flow and volume of the suspension can easily be seen during the procedure and can be regulated by the administrator so that no stress is put onto the cyst by overfilling resulting in the suspension being pushed out of the cyst due to lack of space. Thus, in another aspect of the invention, disclosed are methods of administering a composition comprising antineoplastic particles to an epithelial cyst of a subject, the method comprising injecting the particles using endoscopic ultrasound guided-fine needle injection. This procedure is also applicable to injection into solid tumors such as malignant tumors. In some embodiments, the antineoplastic particles have a mean particle size (number) of from 0.1 microns to 5 microns. In other embodiments, the antineoplastic particles have a mean particle size (number) of from 0.3 microns to 5 microns. In preferred embodiments, the antineoplastic particles are crystalline particles. In some embodiments, the antineoplastic particles are taxane particles. In some embodiments, the taxane particles comprise at least 95% of the taxane. In some embodiments, the taxane particles exclude albumin. In some embodiments, the taxane particles are paclitaxel particles or docetaxel particles.

A non-limiting exemplary procedure for injection of the composition into a pancreatic cyst is as follows: a linear array echoendoscope is inserted via the mouth and advanced to the stomach or duodenum, whichever provides the best access to the cyst. A 22-gauge fine needle aspiration (FNA) needle is luer locked into the accessory channel of the echoendoscope. The needle tip is maintained in the cyst for the duration of the procedure. Using a syringe, the cyst fluid is aspirated from the cyst (usually up to 80% of the original volume of the cyst, but more than 80% of the cyst fluid can be aspirated). The volume of cyst fluid withdrawn is determined. The needle is then filled with the composition, and is injected directly into the cyst. The volume of the composition injected into the cyst can be at a volume equal to the volume of cyst fluid aspirated.

The methods can further comprise additional treatments such as cyst fluid aspiration by endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) performed before the intracystic injection procedure as stated above.

The epithelial cyst is successfully treated when the cyst is reduced in volume/size, has reduced growth rate, is eliminated, or is ablated after treatment by intracystic injection of the composition.

EXAMPLES

The present invention will be described in greater detail by way of specific examples. The following examples are offered for illustrative purposes only, and are not intended to limit the invention in any manner. Those of skill in the art will readily recognize a variety of noncritical parameters, which can be changed or modified to yield essentially the same results.

Example 1. Production of Paclitaxel Particles and Docetaxel Particles Materials and Methods

Raw paclitaxel and docetaxel were purchased from Phyton Biotech (British Columbia, Canada), lot number FP2-15004 and DT7-14025, respectively. Both were characterized in their raw form. The milling of both drugs was accomplished using a Deco-PBM-V-0.41 mill (Deco). The milling conditions for both compounds were as follows:

-   -   Ball size=5 mm     -   RPM=600     -   Processing time=6 min     -   Room temperature.

Preparation of Paclitaxel Particles

A solution of 65 mg/mL of paclitaxel was prepared in acetone. A BETE MicroWhirl® fog nozzle (BETE Fog Nozzle, Inc) and a sonic probe (Qsonica, model number Q700) were positioned in the crystallization chamber approximately 8 mm apart. A stainless steel mesh filter with approximately 100 nm holes was attached to the crystallization chamber to collect the precipitated paclitaxel nanoparticles. The supercritical carbon dioxide was placed in the crystallization chamber of the manufacturing equipment and brought to approximately 1200 psi at about 38° C. and a flow rate of 24 kg/hour. The sonic probe was adjusted to 60% of total output power at a frequency of 20 kHz. The acetone solution containing the paclitaxel was pumped through the nozzle at a flow rate of 4.5 mL/minute for approximately 36 hours. Paclitaxel nanoparticles produced had an average number-weighted mean size of 0.81 μm with an average standard deviation of 0.74 μm over three separate runs.

Preparation of Docetaxel Particles

A solution of 79.32 mg/mL of docetaxel was prepared in ethanol. The nozzle and a sonic probe were positioned in the pressurizable chamber approximately 9 mm apart. A stainless steel mesh filter with approximately 100 nm holes was attached to the pressurizable chamber to collect the precipitated docetaxel nanoparticles. The supercritical carbon dioxide was placed in the pressurizable chamber the manufacturing equipment and brought to approximately 1200 psi at about 38° C. and a flow rate of 68 slpm. The sonic probe was adjusted to 60% of total output power at a frequency of 20 kHz. The ethanol solution containing the docetaxel was pumped through the nozzle at a flow rate of 2 mL/minute for approximately 95 minutes). The precipitated docetaxel agglomerates and particles were then collected from the supercritical carbon dioxide as the mixture is pumped through the stainless steel mesh filter. The filter containing the nanoparticles of docetaxel was opened and the resulting product was collected from the filter.

Docetaxel nanoparticles produced had an average number-weighted mean size of 0.82 μm with an average standard deviation of 0.66 μm over three separate ethanol runs.

Particle Size Analysis

Particle size was analyzed by both light obscuration and laser diffraction methods. An Particle Sizing Systems AccuSizer 780 SIS system was used for the light obscuration method and Shimadzu SALD-7101 was used for the laser diffraction method. Paclitaxel nanoparticles were analyzed using 0.10% (w/v) sodium dodecyl sulfate (SDS) in water as the dispersant. Docetaxel nanoparticles were analyzed using isopar G as the dispersant.

Paclitaxel suspensions were prepared by adding approximately 7 mL of filtered dispersant to a glass vial containing approximately 4 mg of paclitaxel particles. The vials were vortexed for approximately 10 seconds and then sonicated in a sonic bath approximately 1 minute. If the sample was already suspended, 1:1 solution of paclitaxel suspension to 0.1% SDS solution was made, vortexed for 10 seconds, and sonicated in the sonic bath for 1 minute.

Docetaxel suspensions were prepared by adding approximately 7 mL of filtered dispersant to a plastic vial containing approximately 4 mg of docetaxel particles. The vial was vortexed for approximately 10 seconds and then sonicated in a sonic bath for approximately 2 minutes. This suspension was used for laser diffraction analysis. Unused suspension was poured into a 125 mL particle-free plastic bottle, which was then filled to approximately 100 mL with filtered dispersant. The suspension was vortex for approximately 10 seconds and then sonicated in the sonic bath for approximately 2 minutes. This diluted suspension was used for light obscuration analysis.

A background test was first performed prior to analyzing particles on the AccuSizer 780 SIS. A new particle-free plastic bottle was filled with blank suspension solution by pumping from a reservoir, using a peristaltic pump, through a 0.22 μm Millipore filter and into the bottle. A background analysis was run to ensure the particle/mL count was below 100 particles/mL. A small amount of paclitaxel suspension, 5-100 μL, depending upon concentration of solution, was pipetted into the plastic bottle in place from the background test and was filled with ˜100 mL dispersant and the analysis was started. Counts were monitored and paclitaxel solution added to reach and/or maintain 6000-8000 particle counts/mL during the entire analysis. Once the analysis was completed, the background data was removed and any measurement with less than four counts was removed.

To analyze particles on SALD-7101 using a batch cell, the analysis was started by choosing Manual Measurement. The refractive index was set as 1.5 to 1.7. The batch cell was filled with filtered dispersant just past the etched line. The blank measurement was ran. A small amount of API (paclitaxel or docetaxel) suspension was pipetted, generally <1 mL, depending upon concentration of solution as low as 100 μL, into the batch cell as needed to achieve an acceptable absorbance between 0.15 and 0.2 absorbance unit. The measurements were executed, and the resulting graph with the highest level of confidence was selected; background was automatically accounted for.

BET Analysis

A known mass between 200 and 300 mg of the analyte was added to a 30 mL sample tube. The loaded tube was then mounted to a Porous Materials Inc. SORPTOMETER®, model BET-202A. The automated test was then carried out using the BETWIN® software package and the surface area of each sample was subsequently calculated.

Bulk Density Analyte

Paclitaxel or docetaxel particle preparations were added to a 10 mL tared graduated cylinder through a plastic weigh funnel at room temperature. The mass of the drug was measured to a nearest 0.1 mg, the volume was determined to the nearest 0.1 mL and the density calculated.

Dissolution Studies Paclitaxel

Approximately 50 mg of material (i.e.: raw paclitaxel, milled paclitaxel, or paclitaxel particles) were coated on approximately 1.5 grams of 1 mm glass beads by tumbling the material and beads in a vial for approximately 1 hour. Beads were transferred to a stainless steel mesh container and placed in the dissolution bath containing methanol/water 50/50 (v/v) media at 37° C., pH 7, and a USP Apparatus II (Paddle), operating at 75 rpm. At 10, 20, 30, 60, and 90 minutes, a 5 mL aliquot was removed, filtered through a 0.22 μm filter and analyzed on a U(V/V) is spectrophotometer at 227 nm. Absorbance values of the samples were compared to those of standard solutions prepared in dissolution media to determine the amount of material dissolved.

Docetaxel

Approximately 50 mg of material (i.e.: raw docetaxel, milled docetaxel, or docetaxel particles) was placed directly in the dissolution bath containing methanol/water 15/85 (v/v) media at 37° C., pH 7, and a USP Apparatus II (Paddle), operating at 75 rpm. At 5, 15, 30, 60, 120 and 225 minutes, a 5 mL aliquot was removed, filtered through a 0.22 μm filter, and analyzed on a UV/VIS spectrophotometer at 232 nm. Absorbance values of the samples were compared to those of standard solutions prepared in dissolution media to determine the amount of material dissolved.

Results

The BET surface area of particles produced using the above protocol and variations thereof (i.e.: modifying nozzles, filters, sonic energy sources, flow rates, etc.) ranged between 22 and 39 m²/g. By comparison, the BET surface area of raw paclitaxel was measured at 7.25 m²/g (FIG. 2 ), while paclitaxel particles made according to the methods of U.S. Pat. Nos. 5,833,891 and 5,874,029 ranged from 11.3 to 15.58 m²/g. Exemplary particle sizes produced using the methods of the invention are shown in Table 1.

TABLE 1 Surface Mean Size St Dev area μm μm m²/g Number Volume Number Volume 1 38.52 0.848 1.600 0.667 0.587 2 33.82 0.754 0.988 0.536 0.486 3 35.90 0.777 1.259 0.483 0.554 4 31.70 0.736 0.953 0.470 0.466 5 32.59 0.675 0.843 0.290 0.381 6 38.22 0.666 0.649 0.344 0.325 7 30.02 0.670 0.588 0.339 0.315 8 31.16 0.672 0.862 0.217 0.459 9 23.90 0.857 1.560 0.494 0.541 10 22.27 0.857 1.560 0.494 0.541 11 26.19 0.861 1.561 0.465 0.546

Comparative studies on bulk density, SSA, and dissolution rates (carried out as noted above) for raw drug, milled drug particles, and drug particles produced by the methods a the present invention are provided in Tables 2 and 3 below. The full dissolution time course for the paclitaxel and docetaxel materials are provided in Tables 4 and 5, respectively.

TABLE 2 Compound: Paclitaxel Raw Particles Characteristic Material Batch 1 Batch 2 Mean Milled Number Mean (um) 1.16 0.83 0.67 0.75 0.89 Volume Mean (um) 1.29 1.42 0.57 1.00 1.35 Bulk Density (g/cm³) 0.26 0.060 0.11 0.085 0.31 Surface Area (m²/g) 10.4 35.6 39.8 37.7 15.0 Dissolution (30 min) 18% 42% 52% 47% 32%

TABLE 3 Compound: Docetaxel Raw Particles Characteristic Material Batch I Batch II Mean Milled Number Mean (um) 1.58 0.92 0.80 0.86 1.11 Volume Mean (um) 5.05 4.88 4.03 4.46 3.73 Bulk Density (g/cm³) 0.24 0.062 0.096 0.079 0.44 Surface Area (m²/g) 15.9 43.0 45.4 44.2 15.2 Dissolution (30 min) 11% 27% 27% 27% 9%

TABLE 4 Paclitaxel Dissolution time course Timepoint Paclitaxel Paclitaxel Milled (minutes) Raw Material Particles Paclitaxel 0 0.0% 0.0% 0.0% 10 14.0% 40.2% 23.0% 20 17.8% 47.6% 30.0% 30 18.4% 51.9% 32.3% 60 23.9% 58.3% 38.6% 90 28.6% 62.9% 43.5%

TABLE 5 Docetaxel Dissolution time course Timepoint Docetaxel Docetaxel Milled (minutes) Raw Material Particles Docetaxel 0 0.0% 0.0% 0.0% 5 3.2% 12.1% 3.2% 15 6.9% 21.7% 5.9% 30 11.2% 27.2% 9.3% 60 16.4% 32.9% 12.2% 120 22.4% 38.9% 13.6% 225 26.8% 43.1% 16.0%

Example 2. Phase 2 Study—Intracystic Injection of Paclitaxel Particles in Subjects with Mutinous Cystic Pancreatic Neoplasms Objectives and Purpose of the Study

The primary objective of this study was to evaluate the safety and tolerability of paclitaxel particles (referred to as NANOPAC®) injected directly into mucinous cystic pancreatic neoplasms by endoscopic ultrasound-guided fine needle injection (EUS-FNI). Secondary objectives were (a) to describe the PK of NANOPAC when administered into the cyst within the pancreas, and (b) to determine whether any of the NANOPAC dose concentrations (6, 10, to 15 mg/mL) show signs of preliminary efficacy.

Description of the Study Design

In this open-label trial, up to 30 subjects with mucinous cystic pancreatic neoplasms underwent endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) as part of Standard of Care (SOC). Once there was a diagnosis and confirmation of mucinous cystic pancreatic neoplasm, subjects received intracystic NANOPAC via EUS-FNI. Subjects were followed for cyst response to therapy (as shown by imaging) and concentration of paclitaxel in the systemic circulation post-injection (as determined by PK analysis).

Subjects were enrolled in sequential, escalating cohorts of NANOPAC at concentrations of 6, 10, or 15 mg/mL injected directly into the cyst within the pancreas at a volume equivalent to the amount of fluid removed from the cyst. The study included a dose escalation phase and a dose confirmation phase.

Cyst Volume Calculations

If more than one cyst was present in the pancreas of a subject, the Investigator selected a single target cyst and treated only this target cyst. The single target cyst must be a diameter of at least 2 cm but no more than 4 cm; the diameter was measured at the widest point of the cyst. Imaging with magnetic resonance cholangiopancreatography (MRCP), CT scan, or fluorodeoxyglucose-positron emission tomography (FDG-PET) was used to visualize and measure the cyst to confirm subject eligibility and to estimate cyst volume. The same imaging modality used prior to enrollment was repeated at the 3-month and 6-month timepoints. The exact cyst volume calculation was based on measurements performed with endoscopic ultrasound during the NANOPAC injection procedure. Injection volume will be based on, and equal to, the amount of fluid removed from the cyst.

Dose Escalation of Cohorts

Cohorts were enrolled sequentially starting at the lowest dose (6 mg/mL). Each cohort had a planned minimum of three subjects. All data from the first three subjects in a cohort was reviewed and evaluated to determine whether the dose received is considered safe and tolerable, and to determine if dose escalation may occur. The data on the three subjects was reviewed once they had completed the two-week follow-up visit, and the safety and tolerability were assessed, which included reference to dose-limiting toxicities (DLT). A determination was made whether to: (a) escalate to the next dose level cohort (no DLT); (b) add three additional subjects to the current cohort (one DLT); or (c) return to the previous (lower) dose cohort and expand by three subjects (more than one DLT).

The dose most suitable for further evaluation was the highest dose with an acceptable safety and tolerability profile. If one or fewer subjects in a six-subject cohort, or no subjects in a three-subject cohort at the highest dose, experienced a DLT, that cohort was taken into the dose confirmation phase. If greater than one subject in a six-subject cohort experienced a DLT, the previous dose was taken into the dose confirmation phase.

Dose Confirmation Phase

Once the dose deemed appropriate for expansion and further evaluation was determined, additional subjects were enrolled to provide up to a total of 12 subjects dosed at that dose level.

PK Analysis

Plasma samples were taken on the day of injection prior to injection and at 1 and 2 hours after NANOPAC injection, as well as at all other study visits, to characterize the PK of intracystic NANOPAC. Subjects were followed for six months after NANOPAC injection for safety and response to therapy. A further follow-up was conducted at six months.

NANOPAC Study Agent

Paclitaxel particles (referred to as NANOPAC) were presented as a sterilized white powder in a sealed vial within a study kit. The mean particle size (number) of the used in this study was 0.793 microns, the SSA was 29.9 m²/g, the bulk density (not tapped) was 0.0877 g/cm³, and the assay was approximately 100% paclitaxel.

The study agent for all treatment groups was in kits containing: one 60 cc vial of NANOPAC powder, 306 mg per vial, sterile; one vial of Sterile Reconstitution Solution (1% Polysorbate 80, NF in normal saline solution (0.9% Sodium Chloride for Injection, USP)), 7 mL per vial, as a carrier; and one instructions for use (IFU) insert with instructions for reconstitution of the NANOPAC powder in the Sterile Reconstitution Solution to form a suspension and subsequent dilution of the suspension to the required dose (6 mg/mL, 10 mg/mL, or 15 mg/mL) with a diluent. A diluent of normal saline solution (0.9% Sodium Chloride for Injection, USP) will be used. The IFU additionally had instructions for the dose withdrawal procedure. The kits were placed in a carton. The kits were provided for a once-only use and will be assigned to one subject only.

Preparation of the NANOPAC Suspension

The NANOPAC suspension was prepared using the materials supplied in the study agent kit (i.e., one 60 cc vial of NANOPAC powder, 306 mg per vial, sterile; and one vial of Sterile Reconstitution Solution (1% Polysorbate 80, NF in 0.9% Sodium Chloride for Injection, USP), 7 mL per vial, as a carrier) plus a diluent of normal saline solution (0.9% Sodium Chloride for Injection, USP), following the procedures below to prepare 3 different dose level suspensions (6 mg/mL, 10 mg/m, and 15 mg/mL):

6 mg/mL NANOPAC in 0.1% Polysorbate 80/0.9% Saline:

-   -   1. Using a syringe with an 18-gauge needle or larger, added 5.0         mL of the sterile 1% polysorbate 80 reconstitution solution into         the 60 cc NANOPAC powder vial.     -   2. Vigorously hand shook with inversions to make sure all the         particles adhering to the interior of the vial and stopper are         wetted.     -   3. Continued shaking for 1 minute and examined the suspension         for any large clumps of particles.     -   4. Immediately after shaking, used a syringe with an 18-gauge         needle or larger to add 46 mL of 0.9% sodium chloride for         injection, USP to the vial and hand shook the vial for another 1         minute. Periodically examined the suspension for any large         visible clumps. If present, continued hand mixing until the         suspension is properly dispersed.     -   5. After mixing, allowed the suspension to sit undisturbed for         at least 5 minutes to reduce entrapped air and foam.         10 mg/mL NANOPAC in 0.16% Polysorbate 80/0.9% Saline:     -   1. Using a syringe with an 18-gauge needle or larger, added 5.0         mL of the sterile 1% polysorbate 80 reconstitution solution into         the 60 cc NANOPAC powder vial.     -   2. Vigorously hand shook with inversions to make sure all the         particles adhering to the interior of the vial and stopper are         wetted.     -   3. Continued shaking for 1 minute and examined the suspension         for any large clumps of particles.     -   4. Immediately after shaking, used a syringe with an 18-gauge         needle or larger to add 25.6 mL of 0.9% sodium chloride for         injection, USP to the vial and hand shook the vial for another 1         minute. Periodically examined the suspension for any large         visible clumps. If present, continued hand mixing until the         suspension is properly dispersed.     -   5. After mixing, allowed the suspension to sit undisturbed for         at least 5 minutes to reduce entrapped air and foam.         15 mg/mL NANOPAC in 0.25% Polysorbate 80/0.9% Saline:     -   1. Using a syringe with an 18-gauge needle or larger, added 5.0         mL of the sterile 1% polysorbate 80 reconstitution solution into         the 60 cc NANOPAC powder vial.     -   2. Vigorously hand shook with inversions to make sure all the         particles adhering to the interior of the vial and stopper are         wetted.     -   3. Continued shaking for 1 minute and examined the suspension         for any large clumps of particles.     -   4. Immediately after shaking, used a syringe with an 18-gauge         needle or larger to add 15.4 mL of 0.9% sodium chloride for         injection, USP to the vial and hand shook the vial for another 1         minute. Periodically examined the suspension for any large         visible clumps. If present, continued hand mixing until the         suspension is properly dispersed.     -   5. After mixing, allowed the suspension to sit undisturbed for         at least 5 minutes to reduce entrapped air and foam.

After the NANOPAC suspension was fully reconstituted (including final dilution) to the required dose (6, 10, or 15 mg/mL, according to cohort assignment), and at the time of dose administration, a suitable volume for use (based on the cyst size) was withdrawn from the vial into a syringe for use in the administration to the subject.

Dosing and Administration: Intracystic Administration Procedure by Endoscopic Ultrasound-Guided Fine Needle Injection (EUS-FNI)

On the day of NANOPAC administration, the subject received parenteral antibiotic prophylaxis. The subject was positioned in the left lateral decubitus position and was sedated by an anesthesiologist or delegate using monitored anesthesia care (MAC) with or without airway intubation. A linear array echoendoscope was inserted via the mouth and advanced to the stomach or duodenum, whichever provided the best access to the cyst. The cyst was measured using electronic calipers and the size recorded.

The stylet was removed from a 22-gauge fine needle aspiration (FNA) needle and the needle was luer locked into the accessory channel of the echoendoscope. Doppler ultrasound imaging was used to verify lack of intervening vascular structures in the path to the cyst. The needle tip was maintained in the cyst for the duration of the procedure. Using a syringe, the Investigator used their discretion to aspirate the cyst fluid (usually up to 80% of the original volume of the cyst). The volume withdrawn was documented in the source, and a sample was sent for analysis. The needle was then filled with the fully reconstituted NANOPAC suspension from the syringe prepared previously and was injected directly into the cyst at a volume equal to the volume of cyst fluid aspirated. NANOPAC was injected directly into the cyst within the pancreas using endoscope ultrasound guidance.

Starting Dose and Escalation Schedule

NANOPAC was administered in concentrations based on cohort assignment. Investigators administered a volume equal to the volume of cyst fluid aspirated. The first cohort received 6 mg/mL NANOPAC; the second cohort received 10 mg/mL NANOPAC; and the third cohort received 15 mg/mL NANOPAC.

Cohorts were enrolled sequentially starting at the lowest dose (6 mg/mL). Each cohort had a planned minimum of three subjects, each receiving a single dose of the study agent. Escalation to the next cohort proceeded following review of data. Data from the three subjects in a cohort, including all DLT described in this section, was reviewed and evaluated to determine if the dose received was considered safe and tolerable, and to determine if dose escalation may occur. The data on the three subjects was reviewed once they had completed the two-week follow-up visit and assessed safety and tolerability. It was determined whether to: (a) escalate to the next dose level cohort (no DLT); (b) add three additional subjects to the current cohort (one DLT); or (c) return to the previous (lower) dose cohort and expand by three subjects (greater than one DLT). If one or fewer subjects in a six-subject cohort, or no subjects in a three-subject cohort at the highest dose, experienced DLT, that cohort was taken into the Dose Confirmation Phase. If greater than one subject in a six-subject cohort experienced DLT, the previous dose was taken into the Dose Confirmation Phase. A single administration of NANOPAC was injected directly into the cyst within the pancreas.

Endpoints

The primary endpoint was safety and tolerability, as assessed by adverse event (AE), changes in vital signs, laboratory results, and physical examination at one month following NANOPAC injection. Safety and tolerability was continued to be assessed until the six-month end-of-study visit. The secondary endpoints were (a) Concentration of paclitaxel in the systemic circulation post-injection (as determined by PK analysis), and (b) Cyst volume response (imaging).

Preliminary Results

Subject 04001: 69-year old woman with a pancreatic cyst of the uncinate process. At time of injection, the widest diameter of the cyst was 2.8 cm. 3.5 mL of 6 mg/mL NanoPac suspension was injected into the cyst to match the quantity of cyst fluid aspirated. At 3-month follow-up, CT scan revealed the cyst had stabilized with the widest diameter remaining at 2.8 cm. At 6-month follow-up, imaging revealed a 3.1×2.3×2.3 cm cyst with a decrease in volume from 10.1 cc to 8.5 cc. No ductal dilation or new focal masses were observed.

Subject 04002: 72-year old woman with a pancreatic cyst. At time of injection, the maximum diameter of the cyst was 2.5 cm. The infection procedure demonstrated that the cyst remained at 2.5 cm, from which 3 mL of cyst fluid was aspirated and 2.5 mL of 6 mg/mL (15 mg total) NanoPac suspension was injected.

Subject 04003: 55-year old woman with a pancreatic cyst at the pancreatic head. A EUS-FNA was performed revealing a cyst of the pancreatic head with a maximum diameter of 2.0 cm, from which 3 mL of cyst fluid was aspirated. 3-months later, the maximum diameter of the cyst had increased from 2.0 cm to 2.7 cm at which time 4 mL of cyst fluid was aspirated and an equal volume of NanoPac suspension (at 6 mg/mL therefore 24 mg total) was injected. At 3-month follow-up, imaging via MRI demonstrated a cyst with a 2.1 cm maximum diameter. Thus, the cyst diameter reduced from 2.7 cm to 2.1 cm in 3 months after the injection of the NanoPac suspension.

Example 3. Intermediate-Size Patient Population Expanded Access of NanoPac® for the Treatment of Mucinous Cystic Pancreatic Neoplasms

In this intermediate-size patient population expanded access protocol, up to ten subjects with mucinous cystic pancreatic neoplasms will receive NanoPac suspension injection into the pancreatic cyst following fine needle aspiration, in a volume equal to the volume of fluid removed. Injection will be via endoscopic ultrasound (EUS) following fine needle aspiration of the cyst fluid.

NanoPac (sterile nanoparticulate paclitaxel) powder for suspension is presented as a white powder, 306 mg, provided in a sealed vial within a study kit. The kit will also include one vial of Sterile Reconstitution Solution (1% Polysorbate 80, NF in normal saline solution (0.9% Sodium Chloride for Injection, USP)), and one pre-printed Instructions for Use (IFU) insert in a 2 ct kit. The suspension will be prepared and diluted with normal saline solution and/or lactated Ringer's solution to a concentration of 10 mg/mL suspension and placed into a syringe.

On the day of NanoPac administration, the subject will receive parenteral antibiotic prophylaxis. The subject will be positioned in the left lateral decubitus position and will be sedated by an anesthesiologist or delegate using monitored anesthesia care (MAC) with or without airway intubation. A linear array echoendoscope will be inserted via the mouth and advanced to the stomach or duodenum, whichever provides the best access to the cyst. The cyst will be measured using electronic calipers and the size recorded.

The stylet will be removed from a 22-gauge fine needle aspiration (FNA) needle and the needle will be luer locked into the accessory channel of the echoendoscope. Doppler ultrasound imaging will be used to verify lack of intervening vascular structures in the path to the cyst. The needle tip will be maintained in the cyst for the duration of the procedure. Using a syringe, the Investigator will use their discretion to aspirate the cyst fluid (usually up to 80% of the original volume of the cyst). The volume withdrawn will be documented in the source, and a sample will be sent for analysis. The needle will then be filled with the study treatment, NanoPac, from the syringe, and NanoPac will be injected at a volume equal to the volume of cyst fluid aspirated.

During the immediate follow-up period (first two weeks) review of safety and possible paclitaxel-related toxicities will be evaluated, and further follow up will be as per standard of care and will include imaging at 3 and 6 months post NanoPac injection.

Example 4. In-Vitro Release Testing Study. Comparative Measurements of Paclitaxel and Docetaxel Concentration Equilibration Across Natural and Synthetic Membranes

An in-vitro release testing study was conducted to comparatively measure the flux of formulations of various forms of paclitaxel and docetaxel across natural epithelial membranes.

Test Articles

-   -   NanoPac (nanoparticulate paclitaxel powder, approximately 98%         paclitaxel with a mean particle size (number) of 0.827 microns,         a SSA of 27.9 mg²/g, and a bulk density (not tapped) of 0.0805         g/cm³ used in this example) in Suspension, 6 mg/mL.     -   NanoDoce (nanoparticulate docetaxel powder, approximately 99.5%         docetaxel with a mean particle size (number) of 0.915 microns, a         SSA of 33.4 mg²/g, and a bulk density (not tapped) of 0.0675         g/cm³ used in this example) in Suspension, 10 mg/mL.     -   Abraxane® diluted to 6 mg/mL.     -   Paclitaxel solution for injection diluted to 6 mg/mL.     -   Docetaxel solution for injection diluted to 10 mg/mL.

Epithelial membrane substrates: Porcine bladder and porcine intestine were sourced. Upon receipt of the bladder and intestine, the membranes were stored at −20° C. until used. Prior to use, the membranes were removed from the freezer and allowed to thaw fully at ambient temperature.

Equipment

-   -   Franz-type Diffusion Cells (FDCs): 64 diffusion cells with 3.3         ml receptor volume and a 0.55 cm2 receptor fluid exposure         surface area.     -   Stirring Dry Block Heaters: Reacti-Therm #1.8823 stirring dry         block heaters were used to maintain the receptor fluid at         32±0.5° C. with constant stirring throughout the study.     -   Agilent 1260 HPLC unit with a G16120 MS detector, ID #:         TM-EQ-069.

Receptor Fluid: The receptor fluid consisted of 60 vol. %/40 vol % methanol/water at pH 4 with 0.01 wt % NaN₃ (added as a preservative). The solubility of paclitaxel and docetaxel in the Receptor Fluid was determined to be sufficient to maintain sink conditions throughout the study. After mixing and degassing the Receptor Fluid it was filtered through a ZapCap CR 0.2 μm membrane under vacuum; the Receptor Fluid, so filtered, was stirred for an additional 20 minutes under vacuum.

Experimental Procedure

-   -   1. The receptor wells were filled with degassed Receptor Fluid         using a pipette.     -   2. A 6 mm by 3 mm diameter Teflon coated magnetic stir bar was         introduced into each receptor well.     -   3. The defrosted and washed bladder or intestine pieces were         examined and only areas of even thickness and with no visible         surface damage were used.     -   4. The bladder and intestine pieces were cut into approximately         2 cm×2 cm squares using skin scissors. The square sizes were         adjusted as necessary according to the shape and dimensions of         the substrate, but were selected to be approximately uniform in         size among all FDCs.     -   5. Substrate pieces were centered on each inverted donor         compartment.     -   6. The donor and receptor well compartments were then aligned         and clamped together with a pinch clamp, ensuring that the         substrate pieces were centered between both donor and receptor         wells.     -   7. Additional Receptor Fluid was added as necessary. Air bubbles         in the receptor well, if any, were removed by tilting the FDC         assembly such that the air escapes along the sample port.         Receptor wells were filled with approximately 3.3 ml of Receptor         Fluid.     -   8. The assembled FDCs were placed into stirring dry block         heaters which were preheated to 32° C. The Receptor Fluid was         continuously agitated via the magnetic stir bar.     -   9. After 20 minutes, the surface of the membranes in each FDC         was examined. If the membranes appeared wet or showed signs of         being compromised, the cell was discarded.

Test Article Application Procedure: After the membrane integrity tests were complete and the cells appropriately sorted, samples of the test articles were then applied to the surface of the substrate. A one-time dosing regimen was used for this study. For all formulations, 100 μl of the formulation was introduced into the donor cells. The donor cells were then capped for the remainder of the experiment. For NanoPac suspension, Paclitaxel solution for injection, and Abraxane, the amount of paclitaxel drug active corresponded to 0.6 wt % correlating to a dose of 1091 μg/cm². For NanoDoce suspension and docetaxel solution for injection, the amount of docetaxel drug active corresponded to 1.0 wt % correlating to a dose of 1818 μg/cm². “Blank” undosed FDC cells were also set up to test for background signal noise. The background noise measured from these “blank” cells was negligible.

Sampling of Receptor Fluid: Using a graduated Hamilton type injector syringe, a 300 μl aliquot was abstracted from the sampling port of each FDC at each of 1, 3, 8, and 24 and 47 hours. Fresh Receptor Fluid was added to each receptor well to replace the volume of fluid abstracted. Each abstracted aliquot was introduced into a well in a 96-well microtiter plate. Samples were stored in a refrigerator at 4-8° C. prior to MS analysis. Samples were analyzed within 5 days of collection.

Analysis of Sample: The samples abstracted from the receptor wells were then analyzed using a MS method. The concentrations of the Active were assayed and reported in each case. After the MS testing was complete, the samples were analyzed using Chemstation software. The AUCs of the paclitaxel or docetaxel peaks were recorded and converted to μg/ml real values using a calibration curve developed from the calibration standards' AUC values and known concentration values. These μg/ml values were imported into the study results Excel workbook. These concentrations were then multiplied by the receptor volume (3.3 mL) and divided by the surface area of the skin exposed to the receptor fluid (0.55 cm²) for an end cumulative amount in μg/cm². For receptor fluid time points greater than 1 hr, this μg/cm² value was corrected for the sample aliquot volumes which were removed to compensate for the dilution caused by replacing the sample volume with fresh buffer solution. As an example, for the second time point at 3 hrs, the dilution factor (300 μl aliquot/3.3 ml receptor volume or 1/11) is multiplied by the μg/cm² value calculated for the 1 hr time point, the result of which is then added to the μg/cm² concentration which is calculated using the 3 hr AUC value.

Results: The results are shown in FIG. 1 , FIG. 2 , and FIG. 3 .

FIG. 1 is a graph of the flux of paclitaxel (delivered dose of paclitaxel active drug across a porcine bladder membrane over time) from various paclitaxel formulations.

FIG. 2 is a graph of the flux of paclitaxel (delivered dose of paclitaxel active drug across a porcine intestinal membrane over time) from various paclitaxel formulations. Note: flux amounts greater than dose amounts were attributable to evaporation of the receptor fluid.

FIG. 3 is a graph of the flux of docetaxel (delivered dose of docetaxel active drug across a porcine bladder membrane over time) from various docetaxel formulations. Note: the 48-hour timepoint was discarded due to evaporation issues with the receptor fluid samples.

As can be seen in the figures, the NanoPac and NanoDoce formulations had the lowest flux across the membranes as indicated by the least amount of active drug delivered over time through the membranes. These results indicate that NanoPac is retained on one side of an epithelial membrane in greater amounts than Abraxane or paclitaxel solution over time. Also, NanoDoce is retained on one side of an epithelial membrane in greater amounts than docetaxel solution over time. This would suggest that NanoPac when injected into an epithelial cyst would reside within the cyst in greater amounts over time than would Abraxane or paclitaxel solution, and that NanoDoce when injected into an epithelial cyst would reside in the cyst in greater amounts over time than would docetaxel solution.

REFERENCES

-   -   ABRAXANE Package Insert. Celgene Company. Rev July 2015.     -   Allen, P. J., & Brennan, M. F. management of cystic lesions of         the pancreas. Adv Surg. 2007; 41:211-228.     -   Ashgate Handbook of Antineoplastic Agents, Gower Publishing         Limited, 2000.     -   Atar, M. et al., EUS Guided Injection of Albumin Bound         Paclitaxel into Mucinous Pancreatic Cysts. Gastrointestinal         Endoscopy, 81(5), AB555.     -   DeWitt, J., McGreevy, K., Schmidt, C. M., & Brugge, W. R.         EUS-guided ethanol versus saline solution lavage for pancreatic         cysts: a randomized, double-blind study. Gastrointest Endosc.         2009; 70(4):710-723.     -   DeWitt, J. M., Al-Haddad, M., Sherman, S., LeBlanc, J.,         Schmidt, C. M., Sandrasegaran, K., et al. Alterations in cyst         fluid genetics following endoscopic ultrasound-guided pancreatic         cyst ablation with ethanol and paclitaxel. Endosc. 2014;         46(06):457-464.     -   Farrell, J. J. Prevalence, diagnosis and management of         pancreatic cystic neoplasms: current status and future         directions. Gut Liver. 2015; 9(5):571-589.     -   Farrell, J. J., Fernández-del Castillo, C. Pancreatic Cystic         Neoplasms: Management and Unanswered Questions.         Gastroenterology. 2013; 144:1303-1315.     -   Fernández-del Castillo, C., & Warshaw, A. L. Cystic tumors of         the pancreas. Surg Clin N Am. 1995; 75(5):1001.     -   Ferrone, C. R., Correa-Gallego, C., Warshaw, A. L., Brugge, W.         R., Forcione, D. G., Thayer, S. P., et al. Current trends in         pancreatic cystic neoplasms. Arch Surg. 2009; 144(5)448-454.     -   Gómez, V., Takahashi, N., Levy, M. J., McGee, K. P., Jones, A.,         Huang, Y., et al. EUS-guided ethanol lavage does not reliably         ablate pancreatic cystic neoplasms (with video). Gastrointest         endosc. 2016; 83(5):914-920.     -   Greer, J. B., & Ferrone, C. R. Spectrum and classification of         cystic neoplasms of the pancreas. Surg Oncol Clin N Am. 2016;         25(2):339-350.     -   Jani, N., Bani Hani, M., Schulick, R. D., Hruban, R. H., &         Cunningham, S. C. Diagnosis and management of cystic lesions of         the pancreas. Diagn Ther Endosc. 2011:1-10.     -   Kirtane, T., & Bhutani, M. S. EUS for pancreatic cystic         neoplasms: The roadmap to the future is much more than just a         few shades of gray. Asian Pac J Trop Med. 2016; 9(12):1218-1221.     -   McGrath, K. Management of incidental pancreatic cysts: which         guidelines? Endoscopy International Open. 2017; 05:E209-E211.     -   Mills, K. M., Johnson, D. M., Middlebrooks, M., & Burton, G. V.         Possible Drug-Associated Pancreatitis after Paclitaxel-Cremophor         Administration. Pharmacother: J Human Pharmacol Drug Ther. 2000;         20(1):95-97.     -   Moyer, M. T., Dye, C. E., Sharzehi, S., Ancrile, B., Mathew, A.,         McGarrity, T. J., et al. Is alcohol required for effective         pancreatic cyst ablation? The prospective randomized CHARM trial         pilot study. Endosc Int Open, 2016; 4(05):E603-E607.     -   Muthusamy, V. R., Chandrasekhara, V., Acosta, R. D.,         Bruining, D. H., Chathadi, K. V., Eloubeidi, M. A., et al. The         role of endoscopy in the diagnosis and treatment of cystic         pancreatic neoplasms. Gastrointest Endosc, 2016; 83(3)481-488.     -   National Cancer Institute (2016). SEER Stat Fact Sheets:         Pancreas Cancer         http://seer.cancer.gov/statfacts/html/pancreas.html Accessed         Sep. 13, 2016.     -   Oh, H. C., Seo, D. W., Lee, T. Y., Kim, J. Y., Lee, S. S.,         Lee, S. K., et al. New treatment for cystic tumors of the         pancreas: EUS-guided ethanol lavage with paclitaxel injection.         Gastrointest Endosc. 2008; 67(4):636-642.     -   Oh, H. C., Seo, D. W., Kim, S. C., Yu, E., Kim, K., Moon, S. H.,         et al. Septated cystic rumors of the pancreas: is it possible to         treat them by endoscopic ultrasonography-guided intervention?         Scand J Gastroenterol. 2009; 44(2):242-247.     -   Oh, H. C., Seo, D. W., Song, T. J., Moon, S. H., Park, D. H.,         Lee, S. S., et al. Endoscopic ultrasonography-guided ethanol         lavage with paclitaxel injection treats patients with pancreatic         cysts. Gastroenterol. 2011; 140(1):172-179.     -   Pitman, M. B., Lewandrowski, K., Shen, J., Sahani, D., Brugge,         W., & Fernandez-del Castillo, C. Pancreatic cysts. Cancer         Cytopath. 2010; 118(1):1-13.     -   Rheology Modifiers Handbook—Practical Use and Application,         Braun, William Andrew Publishing, 2000.     -   Sarr, M. G., Carpenter, H. A., Prabhakar, L. P., Orchard, T. F.,         Hughes, S., van Heerden, J. A., et al. Clinical and Pathologic         Correlation of 84 Mucinous Cystic Neoplasms of the Pancreas: Can         One Reliably Differentiate Benign From Malignant (or         Premalignant) Neoplasms?. Ann Surg. 2000; 231(2):205-212.     -   Tamirisa, N. P., Parmar, A. D., Vargas, G. M., Mehta, H. B.,         Kilbane, E. M., Hall, B. L., et al. Relative contributions of         complications and failure to rescue on mortality in older         patients undergoing pancreatectomy. Ann Surg. 2016;         263(2):385-391.     -   Tanaka, M. Current best practice and controversies in the follow         up of patients with asymptomatic branch duct IPMN of the         pancreas. HPB 2016, 18, 709-711.     -   Tanaka, M., et al., International consensus guidelines 2012 for         the management of IPMN and MCN of the pancreas. Pancreatology.         12 (2012) 183-197.     -   Tanaka, M., et al., Clinical aspects of intraductal papillary         mucinous neoplasm of the pancreas. J Gastroenterol. 2005;         40:669-675.     -   Tanaka, M., Chari, S., Adsay, V., Fernandez-del Castillo, C.,         Falconi, M., Shimizu, M., et al. International consensus         guidelines for management of intraductal papillary mucinous         neoplasms and mucinous cystic neoplasms of the pancreas.         Pancreatol. 2006; 6(1-2):17-32. 

1.-39. (canceled)
 40. A method for treating an epithelial cyst, the method comprising injecting a composition comprising an effective amount of paclitaxel particles and an aqueous liquid carrier directly into the epithelial cyst, thereby treating the epithelial cyst, wherein the paclitaxel particles comprise at least 95% by weight of paclitaxel, wherein the paclitaxel particles have a specific surface area (SSA) of at least 18 m²/g, wherein the paclitaxel particles have a mean particle size (number) of from 0.1 microns to 5 microns, wherein the composition comprises a suspension of the paclitaxel particles dispersed in the carrier.
 41. The method of claim 40, wherein the aqueous liquid carrier comprises 0.9% saline solution.
 42. The method of claim 40, wherein the aqueous liquid carrier comprises a surfactant.
 43. The method of claim 42, wherein the surfactant is a polysorbate.
 44. The method of claim 43, wherein the polysorbate is polysorbate 80, and wherein the polysorbate 80 is present in the aqueous carrier at a concentration of between about 0.01% v/v and about 1% v/v.
 45. The method of claim 40, wherein the composition further comprises a diluent, wherein the aqueous liquid carrier and the diluent form a mixture, and wherein the composition is a suspension of the paclitaxel particles dispersed in the aqueous liquid carrier/diluent mixture.
 46. The method of claim 45, wherein the diluent comprises 0.9% saline solution.
 47. The method of claim 40, wherein the paclitaxel particles have a bulk density (not tapped) of 0.05 g/cm³ to 0.15 g/cm³.
 48. The method of claim 40, wherein the concentration of the paclitaxel particles in the composition is between about 6 mg/mL and about 15 mg/mL.
 49. The method of claim 40, wherein the paclitaxel particles have a mean particle size (number) of from 0.1 microns to 1.5 microns.
 50. The method of claim 40, wherein the paclitaxel particles are crystalline particles.
 51. The method of claim 40, wherein the composition and paclitaxel particles exclude albumin.
 52. The method of claim 40, wherein cyst fluid is withdrawn from the epithelial cyst prior to injecting the composition.
 53. The method of claim 52, wherein the volume of the composition injected into the epithelial cyst is equal to the volume of cyst fluid removed from the epithelial cyst.
 54. The method of claim 40, wherein the epithelial cyst is a pancreatic cyst.
 55. The method of claim 54, wherein the pancreatic cyst is a mucinous pancreatic cyst.
 56. The method of claim 40, wherein the injecting of the composition is conducted by endoscopic ultrasound-guided fine needle injection (EUS-FNI).
 57. The method of claim 40, wherein the epithelial cyst is reduced in volume/size, is eliminated, or is ablated after injection of the composition, and/or wherein pain associated with the epithelial cyst is reduced. 